FINAL REPORT
SYNTHETIC BIOLOGY: PROSPECTIVE PRODUCTS
AND APPLICATIONS FOR FOOD AND FEED –
REQUIREMENTS FOR REGULATION
FS 102068
09/04/2014
Food and Environment Research Agency
Sand Hutton, York
YO41 1LZ
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Project team members

Fera
Dr Villie Flari – project manager
Mr James Blackburn
Dr Qasim Chaudhry
Dr Christine Henry
Mrs Sarah Hugo
Dr Richard Thwaites
External advisor:

Dr Claire Marris, Kings College London
© Crown Copyright [2014]
This report has been produced by the Food and Environment Research Agency under a contract
placed by the Food Standards Agency (the Agency). The views expressed herein are not necessarily
those of the Agency. The Food and Environment Research Agency warrants that all reasonable skill
and care has been used in preparing this report. Notwithstanding this warranty, the Food and
Environment Research Agency shall not be under any liability for loss of profit, business, revenues or
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report.
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SYNTHETIC BIOLOGY: PROSPECTIVE PRODUCTS AND
APPLICATIONS FOR FOOD AND FEED – REQUIREMENTS FOR
REGULATION
Deliverable 3
Fitness for purpose of current regulatory frameworks for Synthetic
Biology food and feed products and applications
1
Contents
Item
1. Introduction and background
2. Representative case studies
3. Evaluation of fitness for purpose of current regulatory
frameworks: case by case study
4. Project key messages - conclusions
5. Recommendations
6. References
7. Annexes
1
Page
4
6
12
29
35
38
44
Image from: https://theconversation.com/inventing-life-patent-law-and-synthetic-biology-5178
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1. Introduction and background
The ultimate aim of the project was to identify (a) potential Synthetic Biology food/feed
products/applications that may come forward for approval in the UK in the foreseeable
future (i.e. in 5 to 10 year time) , and (b) uncertainties and gaps in relation to the relevant
regulatory frameworks that are currently in place to control such products. The project was
realised in two phases to address the above aims (as shown in Figure 1 below). In phase I,
the team developed a step-wise methodology to critically search the publicly available
information with the view to identifying literature items that would potentially indicate
current or imminent Synthetic Biology products and applications in the food and feed
sectors. Thereafter, the project team critically screened the identified literature items and
selected the ones that would be reviewed further and categorised in the next phase of the
project. The results of this phase are provided in detail in Deliverables 12 and 23 of the
project, and are shown in Annexes I-A and I-B respectively.
Phase I
Part 1
Phase I
Part 2
Phase I
Part 3
Phase I
Part 4
Phase I
Part 5
Phase II
• Developed and finalised in Deliverable 1
• Human intelligence - definition of key search strings, and automatic processes
• Produced 811 items
• Developed and finalised in Deliverable 1
• Human intelligence - screening the 811 items in terms of relevance to the working understanding for synthetic biology
products and applications
• Produced 49 items
• Developed and finalised in Deliverable 2
• Human intelligence - further articles were identified following the review of the 49 identified items via discussions of the
• Final enriched inventory included 71 items
• Developed and finalised in Deliverable 2
• Human intelligence - evaluation of the 71 items in terms of how close each one is to the concept of synthetic biology
• Produced 10 items
•
•
•
•
Developed and finalised in Deliverable 2
Human intelligence - detailed assessment of 10 items
Produced 6 items
List of 6 items was enriched via further discussions and produced in total 8 items to consider as representative case studies
• Developed and finalised in Deliverable 3
• Human intelligence – choice of representative case studies to employ in order to evaluate the applicability of current
regulatory frameworks to regulate such products
• Human intelligence - evaluation of fitness for purpose of current regulatory frameworks to regulate synthetic biology food
and feed products and applications
Figure 1: Outline of the work
process in the project. The
work was unfolded in two
phases that reflect the
identification of potential
synthetic biology food and
feed products and
applications (i.e. Phase I) and
the evaluation of the
applicability of current
regulatory frameworks to
regulate such products (i.e.
Phase II).
In phase II, the project team members, in collaboration with FSA, other governmental
departments, and further external experts (e.g. academia), reviewed the current regulatory
frameworks that are considered to be applicable, or potentially applicable, to synthetic
biology, and assessed whether they are sufficient to cover all foreseeable requirements.
2
Deliverable 1: Synopsis of current and projected products/applications of synthetic biology for food/feed in the UK (see
Annex I-A for full text).
3
Deliverable 2: Representative case studies of potential Synthetic Biology food/feed applications and factors influencing
their development and commercialisation (see Annex I-B for full text).
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Phase II was realised via a facilitated one-day meeting that was held at Fera on 18th of
November 20134. The meeting started with an introduction to the topic by the project
manager, followed by splitting the attendees into two breakout groups. Each breakout
group was presented with three case studies to evaluate. One of these case studies was
common to both groups. After 4 hours of structured discussion, the two groups came
together for a review of the discussions on each case study. The variety of expertise
represented by the individuals in the meeting included molecular biology, toxicology, GM,
risk analysis, emerging sciences and technologies, and regulatory frameworks (see Annex II
for further details on the agenda and format of the meeting).
The ultimate aim of the meeting was to provide FSA with a basis to understand the main
safety/regulatory issues and highlight any inadequacies and gaps to address the current and,
most importantly, future-projected Synthetic Biology products/applications in the food/feed
area.
4
The agenda and participants of the meeting are shown in Annex II.
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2. Representative case studies
The case studies for the workshop were selected from the list of representative case studies
that were identified in Phase I – Deliverable 2 (Annex I-B)5.
The fact that potential synthetic biology products share a common theme with genetically
modified food and feed products and applications at this point was acknowledged by the
team and also at the expert facilitated meeting (Figure 2 below).
Figure 2: It is widely recognised that GM and synthetic
biology are based on the same basic technologies (e.g.
genetic modification, genetic engineering). Therefore one
expects that at this point in time (where SynBio potential is
starting to be realised) a large area of potential overlap exists
between GM and SynBio products and applications. A clear
and concise definition or understanding of synthetic biology
will enable the delineation of the boundaries between what
is perceived as a GM or a Synthetic Biology
product/application. The graphical display is for illustration
only, and does not imply any relationship with actual sizes
of GM and/or synthetic biology market size, etc.
Currently, the technological advances in synthetic biology are placed mainly, if not
exclusively, in the area of overlap between GM and synthetic biology; this overlap area also
includes items that most probably look 5-10 year ahead of market realisation.
Project team members prepared a brief profile for each case study to address a number of
dimensions, in particular (a) where and how the product would be manufactured, (b) who
would be exposed and hence potentially at risk, (c) how it would be disposed of, and (d)
whether monitoring and detection of products and the environment would be feasible.
CS 1.1
Coenzyme Q10 synthesised by synthetic biology microbes
Coenzyme Q10 is a natural component of our cellular function that deals with energy
generation in the mitochondria and protects cellular membranes from oxyradical damage. It
5
Following the evaluation of identified items in terms of how close each item is to (a) the concept of synthetic
biology (results shown in Fig. 3 in Deliverable 2 in Annex I-B), and (b) being marketed (results shown in Fig. 4 in
Deliverable 2 in Annex I-B), the project team decided to assess in more detail the items that were mapped closer
to the concept of synthetic biology, i.e. items mapped beyond point “5” in the subjective scale drawn by the
project team members (Fig. 3 in Deliverable 2 in Annex I-B).
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is a potent antioxidant and has many important roles in metabolic, heart and neurological
functions. Deficiency of Co-Q10 can cause certain ailments and hence there is growing
market for the compound as a health/well-being supplement.

How is the product manufactured
The existing products are available in two forms – a limited (and expensive) range produced
as “naturally sourced” products (mainly in the natural trans form through yeast
fermentation), and a vast majority of products based on chemically-synthesised compound
(mixture of cis and trans forms). The synthetic biology concept is to engineer the pathway for
production of Co-Q10 in microorganisms (yeast?; bacteria?) so that there is a greater supply
to meet the increasing market demands for “naturally sourced” trans form of the compound.

Where is it made - UK made consumed and exported or overseas made/imported
and consumed
At proof of concept stage but could be produced in the UK and exported to other countries
or produced in the UK and exported to other countries.

How would the product be used
In: health supplements; nutraceuticals; health-foods6.

Who would use the product
General public; certain consumer groups, e.g. health conscious consumer, elderly, etc.

How it would be disposed
Through normal waste streams, i.e. same routes as the ones for other food waste for existing
consumer products.

Will we be able to detect the presence of the synthetic biology microbe in
consumer products / environment
Certainly detectable as a chemical in consumer products, but may not be detectable in the
environment as it is not likely to be persistent. The main difficulty (perhaps impossibility)
will be in distinguishing the synthetic biology substance from natural sourced substance.
CS 1.2 and CS 2.27 Yeast that can generate vanilla (and potentially other) flavours
in food products in situ
6
Also topical applications, e.g. hand cream, face cream, but this is outside the food/feed remit.
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
How is the product manufactured
Genetic modification of yeast strains used in fermentation of food or drink.

Where is it made - UK made consumed and exported or overseas made/imported
and consumed
Potentially could be made and/or exported from anywhere.

How would the product be used
Live modified yeast added directly to food as a combined raising/fermentation and
flavouring agent.

Who would use the product

Most likely use in industry, by food manufacturers. Also, potentially, direct-toconsumer sales.

How it would be disposed
Depends on application: if industrial fermentation, disposal via industry protocols. If
domestic use is concerned, then disposal through regular food disposal routes is anticipated.
However, if used in uncooked recipes (e.g. fermented drinks), yeast may be consumed live.
There is potential for release via wastewater.

Will we be able to detect the presence of the recombinant yeast in consumer
products / environment
Synthetic biology produced vanillin may not be distinguishable from fully synthetic or
natural vanillin, particularly in prepared foods. However, the detection of genes encoding
vanillin pathway likely to be possible, provided that there is already information

How will the product be labelled?
A crucial issue with respect to vanillin and other food flavourings and additives produced
by synthetic biology will be two questions related with food labelling, especially for
products sold directly to consumers. Will vanillin produced by recombinant yeast be
allowed to be labelled as “natural”? Currently only vanillin produced from plants is labelled
“natural”. Will the product have to be labelled as “GM”? This is to some extent separate
from the safety issue, except that post-market monitoring of unanticipated adverse impacts
cannot be conducted when products are not labelled. In addition, within the EU regulatory
7
Case study 1.2 was addressed by breakout group 1 and was identical with case study 2.2 that was addressed by
breakout group 2. More details on breakout groups and case studies in each group are given in Annex II.
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framework, decisions have to be made by regulatory authorities, for every food and feed
product (including food and feed ingredients such as additives, flavourings and vitamins)
produced by a fermentation process under contained conditions using a GMM, about
whether or not they fall within the scope of EC regulation 1829/2003. A key question is
whether or not GM material is detectable in the final product, and complex distinctions are
made between products made “from” or “with” GMMs. Depending on the answer to these
questions, a product will fall under different regulations, be assessed using different risk
assessment processes, and be subject to different labeling requirements.
These questions are currently being debated by regulatory authorities in the USA and in
Europe with respect to the vanillin produced suing GM yeast; and the answers are not yet
clear. They will, however, have a large effect on the economic success of this product, and
will also determine the social and economic impact on farmers currently growing orchids for
the production of what is currently sold on the market as ‘natural’ vanillin.
CS 1.3
Enhanced levels, and potentially a multitude of supplements
(different vitamins, antioxidants, etc.) by a single strain of synthetic biology
probiotic bacteria

How is the product manufactured
Modification/construction of bacterial strain(s) with probiotic properties, for example:
enhanced functions for persistence in gut and provision of health/nutritional benefits.

Where is it made - UK made consumed and exported or overseas made/imported
and consumed
Potentially could be made and/or exported anywhere.

How would the product be used
Probably as a formulated probiotic culture. Product will be live cells.

Who would use the product
General public: all consumers; medical patients.

How it would be disposed
Live cells would be consumed, so possible route to environment through sewage.

Will we be able to detect the presence of the synthetic biology probiotic bacteria in
consumer products / environment
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Yes, if detectable markers incorporated into strain(s) or if molecular assays are developed to
detect specific modifications.
CS 2.1
Synthetic biology microbes to produce new feedstock compounds
Due to growing health and environmental awareness, there is a continuous demand for
industrial feedstock chemicals that can be derived from ‘natural’ and sustainable sources to
replace the use of synthetic chemicals in a wide range of consumer products (e.g. cosmetics,
food/feed, packaging, detergents, etc.). The synthetic biology concept is to rationally
synthesise/engineer some of the key biosynthetic pathways, e.g. isoprenoid pathway, in
microbes in such a way to obtain specific (also potentially novel) industrially important
chemicals.

How is the product manufactured
Currently, the product is at proof of concept stage.

Where is it made - UK made consumed and exported or overseas made/imported
and consumed
Specific “cell factories” could be developed in the UK and/or the rest of EU; the isolated,
purified products could be used in the UK and/or exported to other countries.

How would the product be used
High value substances will be used by industry to produce consumer goods (cosmetics,
food, packaging, detergents, etc.) that will be used by the common consumer in the normal
way.

Who would use the product
General public: all consumers.

How it would be disposed
Through normal waste streams, i.e. same routes as the ones for other food waste for existing
consumer products.

Will we be able to detect the presence of the synthetic biology microbe in
consumer products / environment
Yes as a chemical, but not as a synthetic biology produced chemical, unless it is novel and is
only produced by a synthetic biology route.
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CS 2.3
Non-leguminous crops able to fix atmospheric nitrogen – reducing
the need for synthetic fertilisers

How is the product manufactured
If it is a crop e.g. cereals/wheat/rice/ seed will be produced in field. Also, possibly in
glasshouses for first bulk up.

Where is it made - UK made consumed and exported or overseas made/imported
and consumed
Seed can be exported from / imported into UK for use.

How would the product be used
Deliberate release would occur in the field. Possibly contained use would occur in nurseries,
and seed production companies.

Who would use the product
Seeds would be used by seed companies, farmers. The crop would be used by food
manufacturers, food technology companies – grain would be used for flour/bread and cakes
production. It could also be used as animal feed, and straw can be used as bedding.

How it would be disposed
Back to soil/ploughed. Product could be put in compost. Also waste can be directed to
anaerobic digestion units.

Will we be able to detect the presence of the genetically engineered crop in
consumer products / environment
Probably as “novel” genes not normally found in cereals – possibly via DNA PCR tests or
protein based test kits.

Additional information
In particular information from (a) JI Gates Project ENSA8, and (b) a particular science group
from the National Institute of Investigation and Technology in Agriculture and Food of
Spain9.
8
https://www.ensa.ac.uk/scientific-background/informing-scientists/our-understanding-of-symbiosis-signalingand-the-tools-available-in-legumes/
9
http://www.cbgp.upm.es/en/nitrogen_fixation.php
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3. Evaluation of fitness for purpose of current regulatory frameworks: case
by case study
During the breakout sessions of the workshop, the participants evaluated the fitness for
purpose of current relevant regulatory frameworks to regulate the products similar to the
ones described in each case study. Initially the groups discussed, and if it was deemed
necessary, enriched the information provided for each case study. Thereafter each group
reviewed relevant regulatory frameworks in terms of their (a) scope and (b) steps,
requirements and procedures to assess each case study against the statutory criteria of the
regulatory frameworks.
A number of existing regulations for Genetically Modified products and applications were
expected to be taken into account, in particular10:

The Directive 2001/18/EC of the European parliament and of the Council on the
deliberate release into the environment of genetically modified organisms and
repealing Council Directive 90/220/EEC11.

The Directive 2009/41/EC of the European Parliament and of the Council on the
contained use of genetically modified micro-organisms12.

The Novel Foods Regulation (Regulation (EC) No 258/97), including possibly the
proposals for its replacement13.
10
Further relevant regulations which were not however discussed are:

Council Directive 2003/61/EC of 18 June 2003 amending Directives 66/401/EEC on the marketing of
fodder plant seed, 66/402/EEC on the marketing of cereal seed, 68/193/EEC on the marketing of material
for the vegetative propagation of the vine, 92/33/EEC on the marketing of vegetable propagating and
planting material, other than seed, 92/34/EEC on the marketing of propagating and planting material of
fruit plants, 98/56/EC on the marketing of propagating material of ornamental plants, 2002/54/EC on the
marketing of beet seed, 2002/55/EC on the marketing of vegetable seed, 2002/56/EC on the marketing of
seed potatoes and 2002/57/EC on the marketing of seed of oil and fibre plants as regards Community
comparative tests and trials.

The council Directive 2002/53/EC of 13 June 2002 on the common catalogue of varieties of agricultural
plant species.
 The council Directive 66/402/EEC of 14 June 1966 on the marketing of cereal seed.
Directive available at: http://eurlex.europa.eu/smartapi/cgi/sga_doc?smartapi!celexapi!prod!CELEXnumdoc&lg=EN&numdoc=32001L0018&mod
el=guichett
12Directive available at: www.biosafety.be/Menu/BiosEur1.html
13Directive available at: http://ec.europa.eu/food/food/biotechnology/novelfood/initiatives_en.htm
11
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
Regulation (EC) No 1829/2003 of the European Parliament and of the Council of
22 September 2003 on genetically modified food and feed14.

Regulation (EC) No 1830/2003 of the European Parliament and of the Council of
22 September 2003 concerning the traceability and labelling of genetically modified
organisms and the traceability of food and feed products produced from genetically
modified organisms and amending Directive 2001/18/EC15.

The Food Additives legislation (i.e. Council Directive 89/107/EEC of 21 December
1988 on the approximation of the laws of the Member States concerning food
additives authorized for use in foodstuffs intended for human consumption)16.

The Flavourings legislation (Council Directive 88/388/EEC of 22 June 1988 on the
approximation of the laws of the Member States relating to flavourings for use in
foodstuffs and to source materials for their production)17.

Regulation (EC) No 1334/2008 of the European Parliament and of the Council of 16
December 2008 on flavourings and certain food ingredients with flavouring
properties for use in and on foods and amending Council Regulation (EEC) No
1601/91, Regulations (EC) No 2232/96 and (EC) No 110/2008 and Directive
2000/13/EC (Text with EEA relevance)18.
The ultimate question for this exercise was to assess whether current regulatory schemes
would be appropriate and sufficient for regulators to assess fully such synthetic biology
products when these would be presented to regulatory authorities for approval. If
participants thought that current regulatory frameworks are not either appropriate and/or
sufficient for this, they were asked to identify the particular gaps in the regulatory
frameworks and disseminate possible ways to address these gaps.
CS1.1
Coenzyme Q10 synthesised by synthetic biology microbes
This is a food supplement and the case study implies that it is produced via a synthetic
biology microorganism. The conventional alternative is sold as a supplement and is
14
Directive available at: http://europa.eu/legislation_summaries/agriculture/food/l21154_en.htm
Directive available at: http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=CELEX:32003R1830:EN:NOT
16Directive available at: http://www.fsai.ie/uploadedfiles/dir89.107.pdf
17Directive available at: http://ec.europa.eu/food/fs/sfp/addit_flavor/flav09_en.pdf
18
Directive available at: http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=CELEX:32008R1334:en:NOT
15
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produced as by-product of yeast fermentation. Currently demand exceeds supply for
“naturally sourced” substance. A number of organisms have been genetically modified to
produce coenzyme Q10 and these organisms could be used for manufacturing.
The desirable product in our case study would be extracted and purified from the synthetic
biology microorganism. This process is a key point, and it indicates that there are two
separate aspects that need to be regulated, in particular:

The process of manufacturing such a product; as the product is purified from the
producing organism before it is sold it is anticipated that there would be an
appropriate product specification of which purity would be the most significant
component.

The actual end-result of the process, i.e. the extracted, purified, product19.
It was concluded that the process followed to manufacture such products would be
adequately regulated by the current GM regulatory frameworks, in particular the Directive
2009/41/EC of the European Parliament and of the Council on the contained use of
genetically modified micro-organisms. The most crucial aspect of this process would be the
choice of the particular microorganism, as this would dictate the particular risk category the
process would be grouped into and the corresponding containment and control measures
required. If the microorganism employed is judged to be classified in category 1, as defined
in the Contained Use Regulations 200020, then no safety issues would be anticipated. If,
however for example an Escherichia coli strain would be used, then safety issues would be
expected.
Products would have particular specifications, and it is anticipated that the emphasis of the
risk assessment and regulatory frameworks would be to assess whether the specifications
are “the same as” the alternative conventional. In this respect this comparability exercise (in
terms of quality characteristics such as purity, activity, safety and efficacy) could be identical
19
It should be emphasised that key issue is whether one is consuming the recombinant organism, e.g. GM crops,
or consuming a chemical made in/by a recombinant organism and subsequently purified, e.g. vanillin. The
manufacture of vanillin in recombinant organisms is covered by existing legislation on contained use and the
safety of the product is covered by existing legislation/ standards.
20
In order for a GMM to be classified in class 1 one should be confident that even in the event of a total breach
of containment the genetically modified organism would be of no or negligible risk to human health or the
environment (as cited in http://www.hse.gov.uk/biosafety/gmo/acgm/acgm31/paper6.pdf) . The CU2000 regs are
intended to protect human health and the environment for GM activities ranging from class 1 to class 4 – the
level of containment increases as the class of activity increases.
14 | P a g e
to the one followed in bio-similarity exercises for different pharmaceuticals21, provided that
there are conventional alternatives to the newly designed synthetic biology product.
There could be a labelling issue depending on whether the coenzyme Q10 was secreted or
had to be extracted from the cell. In the former case, which was considered as unlikely, the
coenzyme Q10 is produced “by GM organisms” whereas in the latter it is “from GM
organisms”22.
The participants agreed that products similar to the ones described in CS 1.1 could be
consumed either directly (e.g. as Q10 supplement tablets) or “indirectly” if, for example, they
would be introduced into a number of different food items in order to enhance aspects of
these food items’ profile. The latter becomes particularly important in cases where
traceability would be very difficult. For this, the country of manufacturing would be very
important. To our knowledge, most probable candidates for manufacturing are USA, EU,
Japan, and China. The questions that regulators would need to ask at this point would be
whether (a) quality controls for such products (such as purity) are up to standard to any of
these countries, and (b) whether there is enough intelligence in place to identify quickly and
reliably the knowledge trails for such products.
The group initially thought once the product of such processes is purified and leaves the
manufacturer then there is no need for labelling (under the current regulatory schemes) as
the final product would be disconnected from the microorganism that produced it.
However, there are a couple of concerns that were voiced and would need to be addressed
by the relevant regulatory frameworks, specifically:

Whether the product could be labelled as natural or not.

Regardless of the answer to the above, whether the product should be labelled as a
synthetic biology one.
Although it may be feasible to verify how similar to a natural alternative a synthetic biology
product is (e.g. via analytical chemistry tools), there may be open questions as to what
constitutes a “natural” product. With regard to the discussion of “natural” flavour, in our
21
USA: http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm290967.html;
EU: http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2013/05/WC500142978.pdf;
http://www.ema.europa.eu/ema/index.jsp?curl=pages/regulation/general/general_content_000408.jsp
22
Details on labelling requirements can be found in Food Standard Agency, UK guidance [Online] Available at:
http://www.food.gov.uk/policyadvice/gm/gm_labelling#ExamplesoflabellingrequirementsunderECRegulationNo.1829/2003forauthorisedGMOs
%28updatedApril2008%29 [Last accessed 17 12 2013]
15 | P a g e
case vanillin, guidance produced by the FSA (produced in 2002 and updated in 2008) has
advice that covers the use of “non-traditional fermentation” etc. as “not natural”23. However,
the issue is much more complex as (a) responsibility for labelling is now split across FSA
and Defra, (b) the legislation is under review and new legislation comes into effect in 2014,
(c) FSA’s guidance on the term “natural” applies to food labelling in general but does not
apply to flavourings as the latter is specifically defined in EU legislation24. Thus, a
flavouring produced by fermentation could qualify as “natural”, even if the fermenting
organism is GM, as the definition does not differentiate between “traditional” and “nontraditional” fermentation. Whether current regulation captures this issue comprehensively,
specifically when considering “true” synthetic biology products, remains an open question.
Inevitably social inputs are very important to the latter issues. Although the Novel Food
regulation is meant to take into account social inputs, it is also understood that peoples’
perceptions and consumer choices are not taken into account in any of the relevant
regulatory frameworks. It is expected that particularly consumer choices should be as
informative as possible, and a number of potential negative views were highlighted during
the breakout group discussions, e.g. playing with nature; effects of processes employed
and/or products on biodiversity; major economic impacts – most probably big earnings
anticipated for large industry25.
In addition to the above, there are two main underlying issues that apply to all of the case
studies assessed. Specifically:

How to keep track of all developments to ensure that regulations are always up to
standards? How much in advance one would need to prepare and how to recognise
the potential gaps / uncertainties early enough?

If problems exist with current regulations concerning food safety (e.g. for current
GM products / applications) then what is the baseline for synthetic biology products?
23
FSA (2008) Criteria for the use of the terms fresh, pure, natural etc. in food labelling [Online:
http://www.food.gov.uk/multimedia/pdfs/markcritguidance.pdf]
24
Directive available at: http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=CELEX:32008R1334:en:NOT
25
Which? (2013) Consumer report. Available online: http://www.staticwhich.co.uk/documents/pdf/the-future-offood---giving-consumers-a-say---which-report-318060.pdf
16 | P a g e
CS 1.2 and CS 2.2
Yeast (i.e. Saccharomyces cerevisiae) that can generate vanilla
(and potentially other) flavours in food products in situ
The case study particularly refers to the production of vanillin and vanillin glucoside26. This
case study was addressed by both breakout groups, in order to identify any major
underlying differences among the two breakout groups. As no major differences between
the thought processes of the groups were identified, the main discussion points in both
groups are presented below in an aggregated form.
It should be noted that if the vanillin was purified from recombinant yeast then it would be
no different from the coenzyme Q10 case study discussed above (CS 1.1).
This case study discusses a process that employs a synthetic biology yeast that could be
designed to manufacture a number of (extracted) products, e.g. flavours in food products
like vanillin, food additives, etc., but could also lead to potential consumption of live yeast
in an array of relevant consumer products27, e.g. beer, yeast’s extract, wine and/or animal
feed products.
Groups considered the question of whether there is equivalence of the synthetic biology
products to the alternative ones, or whether the synthetic biology metabolites are
substantially different, would be fundamental. In this particular example, what is compared
is pure vanillin purified from a recombinant organism versus vanilla pods which contain
vanillin and other subtle flavours. It should be noted that the issue of equivalence from a
deliberate release (DR) regulatory perspective is not necessarily fundamental. However,
applicants will need to evaluate the GM yeast, the newly expressed molecules (in our case
study this would be vanillin) and the product (e.g. bread) for hazard and risk identification
and characterisation. This would include such aspects as toxicology and transgene structure
and stability. The existence of a comparator may help with the risk assessment process.
Participants in group 1 thought that omics could be employed to identify such
equivalence and/or differences.
26
Vanillin production requires the insertion of three genes from different organisms. Vanillin activates the Yap1
gene of yeast and this causes mitochondrial fragmentation. Consequently, an additional gene has been cloned in
yeast that results in the conversion of vanillin to its non-toxic glucoside. Various yeast genes have been
inactivated to stop the formation of vanillin derivatives, e.g. vanillyl alcohol.
27
For example, beer is treated (e.g. sterile filtering) after being brewed but the hypothesis that an alive yeast
could escape sterilisation cannot be rejected.
17 | P a g e
The nature of the process to manufacture such products infers that a synthetic biology
microbe would be deliberately released in the environment28. Although the fermentation
and bulking process would likely take place under GM contained used regulations, the
marketing would likely be regulated under GM food and feed regulations (Directive
1829/200329) rather than the Directive 2001/18/EC. The biggest issue is likely to be getting
approval for deliberate release. To date, no application for marketing a GM microorganism
for food or feed applications has been approved30.
Under the EU regulations, each application would be considered on a case by case basis and
as part of the risk assessment, applicants would need to identify and characterise hazards
and risks associated with the GMM, the newly expressed molecules and the final product.
This would include detailed characterisation of such aspects as toxicology, transgene
structure and stability and any potential adverse environmental effects. There is detailed
guidance on the type of monitoring that would need to be considered. It is hard to envisage
how one could develop an acceptable monitoring plan31 as all types of effects or variables to
be monitored need to be identified, and the tools and systems to measure them should be in
place.
Group 1 thought that in the event such an approval was granted this would be a big
challenge, and they specifically referred to a number of technical difficulties that would
encompass many uncertainties and for which the acquirement of data would be extremely
costly and challenging. In particular:
28
This refers mainly to usage of yeast in brewing and in products in home baking. In other cases, e.g. vanillin,
there will be no deliberate release. The vanillin will be sold as a purified product and not as “flavoured yeast”. If
the spent yeast is used for animal feed it will be killed first in the same way as cells from biopharmaceutical
production.
29
Such products would be placed into ‘Category 4’ according to the classification system for GMM food and feed
products described in the relevant EFSA guidance (http://www.efsa.europa.eu/en/efsajournal/pub/2193.htm).
Under the EU regulations, each application would be considered on a case by case basis and as part of the risk
assessment, applicants would need to identify and characterise hazards and risks associated with the GMM, the
newly expressed molecules and the final product. This would include detailed characterisation of such aspects as
toxicology, transgene structure and stability and any potential adverse environmental effects. There is detailed
guidance on the type of monitoring that would need to be considered.
30
Consents for part B deliberate release of bacteria and viruses for vaccine and clinical trials have been issued by
Defra – see: https://www.gov.uk/genetically-modified-organisms-applications-and-consents [Last accessed 17 12
2013]
31
EFSA (2011) Guidance on the risk assessment of genetically modified microorganisms and their products
intended for food and feed use (http://www.efsa.europa.eu/en/efsajournal/pub/2193.htm). The guidance goes
into some detail regarding what monitoring may be required.
18 | P a g e

The risk assessment and specifically the detection of microbes in the environment
and monitoring of concentration would prove a much more complex procedure,
compared with GM crops, or nanopesticides32. This is mainly due to the fact that
potential survival and reproduction rates of the microbes in diverse environmental
conditions would be largely unknown. The design of experiments to address gaps in
knowledge, in particular its behaviour and fate in the environment, as best as
possible would be a challenge although it should be noted that designing and
undertaking experiments in containment is seen as an essential part of generating
sufficient data to inform applications for deliberate release. Additionally, it is
possible that this information may need to be acquired via structured expert
knowledge elicitation. In the latter case one would need to provide guidelines
regarding devising structured protocols to elicit information, etc. There are two
expert technical committees currently in place in the UK, i.e. (a) the Scientific
Advisory Committee on Genetic Modification33 (contained use), and (b) the Advisory
Committee on Releases into the Environment34 (deliberate release).
Time wise challenges would be included as well, particularly as this is a new
territory so it is likely to be subject to delays. It is anticipated that initially the
European Food Safety Authority would need to assess such products. Whether new,
quicker processes would need to be devised is an open question.
A major regulatory challenge for synthetic biology products would be when there is
no alternative, conventional, equivalent. In such occasion it is hard to see how one
would deal with the requirement for substantial equivalence for risk assessment, e.g.
“as safe as” etc.

Design of the organism which was recognised as the most important dimension in
terms of safety to health and environment35 (where applicable). The robustness of the
synthetic biology microbes and their survival in variable environment conditions, the
competition with native microbes and their survival in different hosts would be
32
In cases where the Environmental Risk Assessment (ERA) deems these as necessary.
See: http://www.hse.gov.uk/aboutus/meetings/committees/sacgmcu/
34
See: https://www.gov.uk/government/organisations/advisory-committee-on-releases-to-the-environment
35
Key aspect would be engineering disabling mutations (e.g. auxotrophic requirements; absence of essential
genes for replication or survival) – this is often referred to as biological containment and limits the ability of the
microorganisms to survive in the wider environment.
33
19 | P a g e
important aspects of relevant risk assessments and consequently of relevant
regulatory frameworks. The group highlighted the fact that, thus far, an assessment
of a genetically modified microbe for deliberate release in food and feed applications
has not happened. However, there are a number of examples of human and
veterinary medical applications (e.g. gene therapy, human & veterinary vaccines),
and the group emphasised that the experience from these could be used to inform
the approach to food products and food applications.
An interesting relevant case concerns the application for a bacterial biosensor
(Escherichia coli) to detect arsenic in ground water36. This case indicates that for safe
organisms there is a regulatory mechanism in place for obtaining exemption from the
contained use directive – this particular application has applied for this exemption. It
was recognised that on this occasion the release of the genetically modified microbe
would be rather a combination of contained and deliberate release (i.e. the
recombinant bacteria would be contained in vials but the application itself takes
place either partially or fully in the field).

Imports of non-labelled or mislabelled products, as monitoring of such inflow to the
UK market would depend on prior information regarding the manufacturing,
description and trade of such products. Any differences among the labelling of
similar products across the different regions (e.g. EU, USA, Japan, China, etc.) may
have an effect on the uptake and consequently the consumption of such products.
Views on whether precise labelling is needed varied among the participants. Group 2
questioned whether any labelling requirements would be any different from the ones
regarding “extreme GM”37. As mentioned above the GM food and feed regulations
define whether or not a product will be subject to specific labelling requirements
indicating that a product is GM. The group also referred to the regulatory
frameworks of other sectors, e.g. cosmetics, where there are no rules for labelling;
36
More information on this application in: http://www.dhaka.diplo.de/Vertretung/dhaka/en/08/Science-andTechnology/Helmholtz__Centre.html, and in Scoping report for “Workshop on Synthetic biology: containment
and release of engineered microorganisms” held on 29 April 2013 at King’s College London:
http://www.kcl.ac.uk/sspp/departments/sshm/research/csynbi/Scoping-Report.pdf
37
The term “extreme GM” was used to describe the insertion of genes for long pathways, although it is
understood that this term would not be adopted officially.
20 | P a g e
flavourings where there are no specific requirements regarding the method of
manufacturing; detergents and packaging: there is no requirement.

Other possible applications of the engineered yeast - as starter yeast for baking or
beer brewing in household settings (e.g. home based baking and/or beer brewing for
personal consumption and/or distribution to friends and family) were considered.
Such applications would fall within the scope of ‘deliberate release’ in the EU’s
regulatory framework because the living engineered yeast will inevitably be released
into the environment. Unless these activities could be closely monitored it would be
impossible to be aware of the extent of deliberate release of synthetic biology
microbes and of potential effects on the environment. In these applications, vanilla
flavour will be added to the bread or beer directly. The fact that the product could be
deliberately released once sold to consumers was recognised by the group as a grey
area that would potentially introduce another layer in the relevant regulatory
frameworks38.
The regulatory authority would monitor any long term health effects following
possible change of consumer behaviour once the “synthetic biology produced and
labelled as natural product” vanilla becomes available.

Consumer views on genetically modified food are known to vary, and in Europe
public opinion indicates that GM food is less acceptable to consumers. GM
regulations do take into account such analyses to address consumer views; for
example, public representations regarding GM releases relate to risks of damage to
the environment, and are not concerned with garnering public attitudes to GMOs..
However, although they do allow for consumer inputs there is a need for greater
clarity over how these are applied in practice. For example, this uncertainty could be
addressed via quantitative risk/benefit and or multi criteria decision making
analyses. Additionally, group 1 considered that lessons learnt on the latter could be
borrowed by regulatory frameworks addressing other sectors, e.g. regulation for
38
From a regulatory perspective this would appear to be key. One might envisage preparation of the food
product containing the live GMO to be undertaken in accordance with the GM contained use regulation.
However, as soon as the live GMO moves out from CU control measures and into a marketing scenario it will
need to be regulated as a Deliberate Release (in particular Part C). If the marketed product is pre-cooked then
presumably there is no longer any live GMO present and it would not need to be regulated under 2001/18/EC.
21 | P a g e
additives39, regulations for medicines40, etc. This point was also very relevant to
concerns that labelling of the product might have an effect on consumer choices; in
particular it may have an impact on a consumer’s inclination to use it. As a result,
long-term cumulative impacts associated with perception of “natural” flavouring
may arise.
Group 1 also referred to a new legislative package that would aim to condense
existing EU food safety laws into five pieces of legislation to facilitate compliance
with controls, inspections and testing, and intends to harmonise the existing
regulatory scheme on official controls, animal health, and control of pests on plants
and plant reproductive material (including seeds)41.

The probability of contamination of animal feed with live synthetic biology yeast was
also considered. On this occasion it was emphasized that the GM regulation allows
for 0.9% contamination42, although it is not clear whether this would be the case for
synthetic biology microorganisms. This may be particularly important as new
sources of feedstock are not regulated, and it is anticipated that industry
requirements would control this (i.e. contractual obligations). Group 2 thought that
on this occasion REACH43 may apply, as feedstocks could be considered as chemical
substances, although currently, no special requirements e.g. for nanomaterials, are
needed.
CS 1.3
Enhanced levels, and potentially a multitude of supplements
(different vitamins, antioxidants, etc.) by a single strain of synthetic biology
probiotic bacteria
The group recognised that several of the risk assessment and regulatory challenges
identified for the case study employing synthetic biology yeast (i.e. CS 1.2 and 2.2) would be
also applicable for this case study. The group also recognised that the risk assessment of
39
For information see: http://ec.europa.eu/food/food/chemicalsafety/additives/new_regul_en.htm
For information on legal framework for medicinal products see: http://ec.europa.eu/health/human-use/legalframework/index_en.htm
41
For more information on this see: http://ehoganlovells.com/cv/3bfca136bbe0c30601c8b8ccac2e10dd20f608e8
42
This labelling exemption applies where the presence of GM is adventitious; in the case of feed for low level
presence of varieties approved in a third country but submitted to the European Food Safety Authority for
assessment.
43
For more information on this see: http://www.hse.gov.uk/reach/
40
22 | P a g e
synthetic biology probiotic bacteria would be even more challenging, as these bacteria
remain alive and form colonies in the gut, whereas yeast dies in the gut environment.
Because of this individuality the group thought one major risk assessment question would
refer to potential horizontal gene transfer. However, the group unanimously agreed that the
major challenge in the risk assessment of such products would relate to the fact that
probiotic bacteria exist in an environment where many very variable factors interact. This
complexity and interspecies variability inevitably brings many challenges, as to how can this
environment be assessed in one way. For example, each individual may carry different
microflora in his/her gut, consumes different food items at different times during the day.
All these potentially create a very different gut environment per each individual.
Another major challenge, not currently addressed by the GM regulatory frameworks, is the
assessment of benefit claims of such products. To our knowledge, the scientific assessments
undertaken by the European Food Safety Authority could not substantiate the health claims
of such products, and are not aimed at doing so; following the EFSA assessments there is no
approval of these products by any of the European Union Member States. A major question
on this would be why one would go down the route of consuming live bacteria instead of
getting the desired nutraceutical via conventional tablets? Although a cell market for this can
be identified (e.g. children; particular consumer groups who cannot take drugs administered
as oral tablets, etc.) the key thing is to think about the concept of such products and
applications in the sector of synthetic biology. For example synthetic biology probiotic
bacteria could secret a compound that could be transformed further, by the gut and/or other
bacteria species in the gut, to a detrimental to health compound. The group agreed that due
to the nature of the product (i.e. live microbes) there are too many potential interactions in
the gut that cannot be foreseen, detected and monitored. This is a major challenge for the
regulation of such products when they are manufactured in a conventional way. The fact
that such products could be manufactured via genetic modifications complicates further
their risk assessment, and consequently, their regulation in terms of health and environment
safety. Earlier points on risk assessment, contained use and biological containment also
apply here. Additionally, the group thought that the consumer acceptability issue of such
products will be interesting, possibly polarising between “yes” and “no way”, but social
23 | P a g e
science scholarship challenges this view and demonstrates that public views tend to be
much more nuanced and ambivalent (Marris, 2001).
Further potential applications of such products refer to animal feed and industrial methane
production. For the former, the group emphasised that possibly more pronounced effects of
feed containing synthetic biology probiotic bacteria may occur, particularly as ruminant
animals rely on gut microflora to digest feed materials. Thus the regulation of such feed
applications would be expected to face a different set of challenges.
The group also questioned whether the labelling of such products is covered by existent
legislation, although it was understood that the challenges for this aspect, and the potential
effects on consumer behaviour, would be similar to the challenges outlined above for CSs 1.1
and 1.2.
The group recognised that regulation requirements for manufacturing such products would
differ across different geographical areas, and deemed it possible that in certain areas less
stringent regulatory requirements than those of e.g. Europe and USA may exist. If such
differences prevail, then these may dictate the countries of manufacturing such products
first. The group emphasised that the manufacturing country however may be different than
the country that would host trials, and thus regulatory fitness for purpose and/or regulatory
challenges could be different for the two processes.
CS 2.1
Synthetic biology microbes to produce new feedstock compounds
This has been on-going for many years and is governed by regulations covering the large scale
culture of recombinant microorganisms. The feedstock chemicals will be purified and sold/
purchased against purity specifications.
The group thought that more detailed information would be needed before evaluating
precisely the fitness for purpose of regulatory frameworks to regulate such products. In
particular, (a) how exactly was the product made, and the molecular composition of the
products concerned, and (b) detailed information on by-products and/or intermediates that
could emerge in the manufacturing process.
A major issue recognised by the breakout group was the need to ensure quality control,
which could have impact/s on risk assessment (i.e. the particular by-products and/or the
intermediates of the process) and/or labelling aspects. The former would be of high interest
24 | P a g e
as any impurities may relate to new biologically active compounds, therefore increasing the
probability of introducing new allergens to the individual and/or the population.
The group thought that such products would be regulated via the Directive 2009/41/EC of
the European Parliament and of the Council on the contained use of genetically modified
micro-organisms, and once more they reiterated that precise information on the product is
needed before evaluating the fitness for purpose of this Directive, e.g. precise definition of
the synthetic biology microbe concerned is required to assess whether the product meets the
definition of GMO, and if not what type of amending is needed.
For Great Britain, the contained use public register provides details of all centres which have
notified their intention to undertaken contained use genetic modification activities; however,
some information is withheld on grounds of national security.
The group thought that it is probable that these products would be manufactured in
places/countries outside the country of application and/or consumption. Thus, one would
need to ensure that the place/country of manufacturing meets the requirements set in the
contained use regulation.
Waste by-products of the process would need to be disposed of safely, and therefore
environmental risks need to be assessed. Whether a life cycle assessment would be fitter for
purpose for the regulation of such products is an open question.
CS 2.3
Non-leguminous crops able to fix atmospheric nitrogen – reducing
the need for synthetic fertilisers
The group thought that information on this product should be described much more
comprehensively before evaluating the fitness for purpose of regulatory frameworks. The
required missing information, listed below, reflects certain uncertainties that should be
addressed via risk and benefit assessments:
 Detailed description of how exactly the product was made44; what is the molecular
composition of the products concerned?
44
To our knowledge the plant is genetically modified to express genes that will enable it to be colonised by
symbiotic rhizobia species. A “dormant” signalling pathway is re-engineered in cereals so that it can recognise
and interact with rhizobial bacteria as the first step in engineering biological nitrogen fixation into cereal crops.
Presumably at some point transformants would need to tested under field conditions but this is not likely to be in
the near future.
25 | P a g e
 Detailed description of how the system works, and in particular where the
genetically modified part/s are expressed, e.g. grain or straw, and how?
 What happens in the roots, and how could the next crop sown in the same field be
affected?
 Would there be any consequences because of overlap with usage of pesticides and
fertilisers?
 If this product would be applied as a rhizobium product, i.e. a bacterium modified
such that it can fix nitrogen in association with a non-leguminous plant, would GM
regulations apply (discussed further below)?
 Demonstration of effectiveness of the product: does it actually produce the desired
effect?
In practice, the desired goal is likely to be achieved by a combination of both of these
options. The first option is covered by existing legislation on GM foods and the second by
regulations covering deliberate release.
The group discussed a number of potentially interesting end products, for example: (a)
products where the genetic modification takes place in the plant only, and the rhizobium is
not modified; the end result would be genetically-engineered plants so that they can interact
with native soil rhizobia; (b) genetically-engineered rhizobia so that they can nodulate nonlegumes; (c) product would be designed to improve efficiency of bacteria; (d) a combination
of (b) and (c). All possible end products fall within the scope of GM Directive for deliberate
release and possibly the GM food and feed regulations; however, in cases (b) and (c) the
group questioned whether the GM food and feed regulations would be applicable because
the plants themselves (which would be destined the food chain) are not modified, they are
simply obtaining nutrients from the modified bacteria. The question then becomes whether
the plants are obtaining additional metabolites that they would not normally be exposed to,
and which could constitute a human or animal health risk.
The developed risk assessment methodologies thus far are designed to assess genetically
modified plants, and have been tested and possibly validated on plants. The processes
addressed by this case study however refer to new methodologies and approaches that
involve the employment and deliberate release of genetically modified bacteria; therefore it
is largely unknown whether the current risk assessment processes would be appropriate to
26 | P a g e
address such new challenges. Particularly as the guidance referring to genetically modified
microorganisms has not been tested to date for deliberate release, e.g. releasing a genetically
modified microbe into soil. Further, additional “fit-for-purpose” risk assessment regarding
potential human health effects may be needed, as the products may be indirectly consumed
by several subpopulation groups.
The group recognised that monitoring the adequacy of any regulatory framework would be
imperative. This may prove to be a further challenge when application of such products
takes place in developing countries.
Although wastes from synthetic biology products are not within FSA’s remit the group
briefly referred to scenarios describing fate of waste, mainly straw. The hypothesis that this
would be recycled via anaerobic digestion was voiced, although the group was unclear as to
what may happen to the genetically modified organisms.
Key points in all group discussion

All experts thought that the GM food and feed regulation would address most of the
challenges involved in the regulation of products like the ones described in the case
studies considered, mainly because the definitions for the case studies would be
covered by the current regulatory schemes. A review of the novel foods regulation
could be an opportunity to address gaps, including issues around labelling and
reporting requirements. However, it was recognised that the speed of authorising or
not the commercialisation of a product would depend on the degree to which the
product is close to “known” rather than unknown.

The examples used in the case studies involved organisms (mostly microorganisms)
that had been subjected to gene manipulation. In some cases the microorganisms had
been modified by the insertion of large DNA segments encoding long biosynthetic
pathways, e.g. for isoprenoid synthesis. As such, the participants agreed that none of
the case studies evaluated was a representative of “true” synthetic biology. In this
context, the participants thought of “true” synthetic biology can be considered as:
o
a cellular organism created from non-cellular components;
o
a new cellular compartment created within a living cell;
o
a new virus created by assembly of genes from unrelated viruses.
27 | P a g e

From all case studies addressed, the experts thought that CS 1.3 (i.e. enhanced levels,
and potentially a multitude of supplements (different vitamins, antioxidants, etc.) by
a single strain of synthetic biology probiotic bacteria) was the closest to “true”
synthetic biology products/applications. Synthetic biology probiotic bacteria are
likely to pose problems for GM regulations: although the regulatory framework is in
place to cater for this work the challenges are in assessing the adequacy of the risk
assessment which is a crucial input to regulatory decision making.

The group thought about possible scenarios that could lead to regulations becoming
inadequate. An example scenario would be to design and create a microorganism
that does not meet the definition of GM in either of these GM regulations. Although
this was considered unlikely as the current definitions are thought to be very broad.
Experts discussed briefly technologically more advanced potential synthetic biology
products, like protocells; however, currently self-replication of such organisms has
not been achieved, therefore the group did not consider these to be within scope.

It was considered unlikely that any “true” synthetic biology organisms would be
involved in food and feed production in the next ten years. Nevertheless, the experts
agreed that when they will be involved in the food and feed production they will
create interesting challenges for risk assessment, labelling and consumer choices and
behaviour.

The legislative requirements for GMOs, in the context of synthetic biology, would
need to address: (a) occupational exposure; (b) environmental adverse effects; (c)
human health adverse effects (indirectly via environmental exposure); (d) labelling
requirements. For GM food and feed (and novel foods) the legislative requirements
would need to address: (a) human health adverse effects; (b) animal health adverse
effects; (c) potential misleading of consumers; (d) nutritional quality; (e) risk
assessment procedures; (f) labelling requirements.
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4. Project key messages - conclusions
Definition of synthetic biology: Setting up clear boundaries between GM
products/applications and synthetic biology products/applications was considered crucial in
order to address the objectives of the project. This however proved to be a major challenge
particularly as “true” synthetic biology products and applications are not identified in the
food and feed sectors. Instead, the group developed a working understanding of what
would constitute a synthetic biology product in food/feed sectors45; the particular working
understanding emphasises the need to address quantitative and qualitative elements of
newly designed synthetic biology products and applications. The latter has been debated
very clearly in a report on ethics in synthetic biology by the European group on Ethics in
Science and New Technologies to the European commission (200946), in particular in section
1.3.1. where the elements of intentional design, synchronisation of techniques, and
complexity of end products appear to contribute critically to the definition of “true”
synthetic biology products and applications47.
45
Working understanding for the Fera / FSA project on synthetic biology products & applications in food and
feed areas comprised two parts:
o Quantitative part: Substantially large synthetic parts of genetic material caused to function in a biological
system.
o Qualitative part: Focus on the engineering aspect of novel, synthetic (i.e. artificial) organisms, and in
particular on “how much of the produced organism was designed as new”.
46
The European Group on Ethics in Science and New Technologies (2009) Ethics in synthetic biology [Online
http://ec.europa.eu/bepa/european-group-ethics/docs/opinion25_en.pdf (accessed 06 12 2013)]
47
1.3.1. To what extent does synthetic biology differ from other existing disciplines?
A key issue to address in synthetic biology is its difference from other disciplines, such as those based on the
insertion of recombinant DNA into organisms. For example, techniques used in synthetic genomics (e.g. the use
of synthetic DNA within an existing cell may be considered to be a recombinant DNA application rather than
synthetic biology). It nevertheless appears that no clear boundary can be drawn between genetic engineering that
is based on recombinant DNA and synthetic biology: the first is the starting point and merges into the second
without a clear cut limit. Nevertheless, recognition of the complexity of biological systems and the intention to
construct an organism with radically new properties may be described as a feature of the new discipline.
Balmer A. and Martin P. have underlined (50) that the word ‘synthetic’ is ambiguous since it can mean either
‘constructed’ or ‘artificial’. The former meaning is preferred by synthetic biologists (BBSRC/EPSRC, 2007), but it
is inevitable that the ‘artificial’ aspect of synthetics is to some extent associated with the term. In fact, attempts
have been made to avoid the word ‘synthetic’ by naming the field ‘constructive biology’ or ‘intentional biology’
(Carlson, 2006), but these terms have not become widely adopted.
The scientific community is still debating whether synthetic biology has introduced a paradigm shift compared
with other biotechnologies. Some have indicated that, in order to distinguish between synthetic biological
fabrications and other approaches, like transgenic organisms, the key difference could be that transgenic
organisms are the result of introducing naturally occurring foreign or mutated DNA (genes) into the organism
(51). Synthetic biology, in contrast, would result in the manufacturing of elements with synthetic raw materials
and with no natural counterpart. (52) Some researchers are producing protocells, that mimic the systems found
in biology but differ in that the DNA contains nucleotides not found in already existing organisms. (53) Synthetic
biology therefore involves the use of standardised parts and follows a formalised design process (Arkin and
Fletcher, 2006). In parallel, synthetic biology involves a different level of sophistication and complexity of the
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Which food and feed subsector will “host” first a synthetic biology product/application: An
important angle to look into potential synthetic biology food and feed products and
applications is to foresee the specific food and feed sub-sectors in which one would expect to
see the first “true” synthetic biology applications commercialised. From all potential
applications reviewed in this project we identified one that appeared to be closest to
commercialisation: the production of artificial vanillin via genetically modified yeast48,49.
Additionally, our project identified one other upcoming synthetic biology food and feed
application albeit in a sector outside the scope of this research50, specifically diagnostics51
(information included in Table 6 in Deliverable 1; Annex I-A). Based on the information
identified and reviewed this far the project team concluded that synthetic biology food and
feed products and applications in the subsectors of (a) flavours and fragrances, and (b)
diagnostics would probably be among the first to appear in the UK market.
Synthetic biology food and feed products and applications and regulations: Our work
identified five case studies to represent potential synthetic biology food and feed products
and applications that are likely to be in the market in 5-10 years; one of these case studies
work done in genetic engineering (where one gene at a time is inserted into an existing biological system),
contrary to synthetic biology, where a whole specialised metabolic unit can be constructed (Stone, 2006 and
Breithaupt, 2006:22).
One novelty that synthetic biology has introduced in the design and use of different bioengineering technological
tools is the notion of intentionality. Synthetic biology uses biotechnology to intentionally design and build
engineered biological systems that process information, manipulate chemicals, fabricate materials and structures,
produce energy, provide food, and maintain and enhance human health and our environment. In parallel,
synthetic biology synchronically uses multiple technologies, such as chemistry, engineering, biology,
information technology and nanotechnology. In that respect, synthetic biology uses technology to manufacture
products that are designed to give rise to knowledge or which serve a given aim, defined by the application area
on which they are built, from bioremedies to ICT, biomedicine, biofuels or biomaterials. What is also distinctive
in synthetic biology is recognition of the complexity of the systems that researchers want to reproduce, the fact
that they work on not just molecular cloning of single genes or gene components as in standard molecular
biology, but on whole interacting genetic networks, genomes and ultimately entire organisms. In this sense, the
results of systems biology, a discipline that studies the relations of different metabolic or developmental
pathways within an organism, are important to synthetic biology.
48
The classification of all reviewed potential synthetic biology food and feed products and applications in terms
of how close each one is (at current time) to commercialisation is described in Deliverable 2 (see figure 4 in
Deliverable 2 – Annex I-B).
49
Evolva SA has constructed a yeast-based fermentation route to both vanillin and other vanilla flavour
components. - See more at: http://www.evolva.com/products/vanilla
50
Biosensors were not included in this research – see deliverable 1 – Annex IA for more information on this.
51
Sample6 technologies (http://sample6.com/) developed the world’s first synthetic-biology based bacteria
diagnostic system capable of enrichment-free detection; see also Annex II in Deliverable 1 for more information
for products of this company.
30 | P a g e
referred to the production of vanillin via an engineered yeast (i.e. CSs 1.2 and 2.2). How fit
are current regulatory frameworks to regulate synthetic biology food and feed products like
the ones selected and reviewed as representative case studies? The regulatory framework
that applies includes GMO contained use (2009/41/EC), the GMO deliberate release
(2001/18/EC), the GMO food & feed regulations, and the Novel foods regulation52. The
general conclusion at the facilitated meeting on the 18th November 2013 was that the
organisms described for each case study appear to meet the legal definition of a GMO or
GMM (genetically modified micro-organism) as defined in European legislation, and that
existing regulations cover in principle the various case studies discussed53. It should be
emphasised that a couple of key issues were identified:

One is whether the recombinant organism is consumed (e.g. as in GM crops,
probiotic bacteria) or whether one is consuming a chemical made in/by a
recombinant organism and subsequently purified (e.g. vanillin, coenzyme Q10). The
manufacture of products in recombinant organisms resulting in an extracted,
purified product is covered by existing legislation on contained use and the safety of
the product is covered by existing legislation/ standards. Regardless, a number of
potential gaps and challenges regarding how the current regulatory schemes could
be applied to regulate such products were identified per each case study. Such gaps
and challenges could either delay substantially or halt the process of approval; thus,
ideally they would need to be addressed in advance of the potential
commercialisation of such products. Major identified gaps and challenges relate to
the following:
o
It was recognised that current assessment of GM is based on existing
comparators, i.e. usually the organism’s conventional counterpart. Due to the
unique design basis of synthetic biology products, and particularly for future
52
For full list of relevant regulatory frameworks see section 3 of the report.
The contained use regulations was seen as a useful vehicle for establishing data to inform the environmental
risk assessment required for marketing. The importance of biological containment was emphasised as a means of
practically limiting the environmental impact of GMOs and would also simplify some of the environmental
considerations. The value of other areas which have involved releasing GMOs (e.g. gene therapy, vaccines) to
inform food and feed applications involving microorganisms was highlighted. Additionally, there are a number
of lessons learnt from regulatory frameworks that are relevant to other sectors, e.g. cosmetics, medicinal
products, etc.
53
31 | P a g e
products, this concept of "substantial equivalence" may not be applicable to
all synthetic biology products and applications.
o
Synthetic biology involves much more complex changes than existing GM
crops, so assessments will be breaking new ground and can be expected to
take considerable time and effort.
o
Novel techniques for inserting new traits are continually being developed,
with recent examples including cisgenesis, intragenesis, epigenetic
modification, oligo-induced mutation induction, zinc finger nuclease induced
mutation, and mutation through homologous recombination. Currently there
is much debate as to whether these new techniques fall under the definition
of genetic modification, as described in Directive 2001/18/EC. If SynBio
organisms are developed using these techniques, then the question as to
whether the GM regulations are applicable to these techniques is of major
relevance.
o
Lack of regulation of “feedstocks” entering the food system (i.e. “feedstocks”
used in manufacture of end products), although quality control is achieved
through contractual arrangements between suppliers and purchasers.
Although this point is not specific to synthetic biology, the fact that synthetic
biology processes are much more complex may introduce further challenges.
o
Need to consider safe use and disposal of the intended product as well as of
the by-products (e.g. anaerobic digestion).
o
Labelling of synthetic biology products: the relevant major questions are (a)
whether consumers would want to (a) differentiate between GM products
and synthetic biology products, and (b) label synthetic biology products?
o
GM soil organisms: is safety of crops included in the assessment under
2001/18?

The approval for any of these products via the current (deliberate release) process for
marketing is a notoriously slow process subject to lack of consensus amongst
member states. To date only two GM events are authorised for cultivation in the
Member States of Europe: maize MON 810, developed by Monsanto to provide
resistance to the European corn borer (Ostrinia nubilialis) and maize T25, developed
32 | P a g e
by Bayer CropScience, which is tolerant to glufosinate ammonium54. A genetically
modified potato EH92-527-1 (“Amflora”), developed for the production of starch for
non-food use, was approved for marketing in 2010 , but the company withdrew it
from the EU market in January 2012. This is related in part to the difficulty in
providing a comprehensive environmental risk assessment and in part due to the
political environment within EU.

Imports of synthetic biology food and feed products and applications would be a
huge challenge for the regulation of such products, as consumption would not be
adequately detected and monitored. 1829/2003 would cover imports of GM food and
feed. Trans boundary movements of ‘living modified organisms’ (which includes
seeds and GMMs) would be captured by the Cartagena Protocol of the Convention
on Biological Diversity.

Last but not least, the value of employing mechanisms to assess the benefits along
with employing the European Food Safety Authority to assess the risks of such
products was voiced.
Overall, the evaluation that took place during this breakout group session indicated that
regulations for upcoming applications should be a well improved version of the current
regulatory schemes.
Novelty of synthetic biology and harmonisation of global regulatory schemes: Overall, the
group identified the uniqueness of the synthetic biology products in that computational
modelling is employed in advance to model new organisms. Pure synthetic biology concepts
refer to designs that never existed before, and these would be an opportunity for novel foods
regulations to be regularly reviewed and updated; the group suggested a frequency of every
5 years, although it is understood that the review of relevant regulatory frameworks should
reflect closely the technical progress of this field. A further challenge, that could cause
further delays, would be reaching agreement between European Union member states on
any reviews. As it was recognised that the production of products like the ones addressed in
54
Of these two maize events, only maize MON810 can be grown in the EU because T25 maize does not have any
varieties registered on the EU Common Catalogue of Varieties.
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CSs 1.1, 1.2, 1.3 would encompass a number of different stages (i.e. manufacturing, trial,
consumption, monitoring, detection) the group emphasised the need of harmonised
regulation across European Union and also potentially worldwide.
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5. Recommendations and next steps
Based on the conclusions reached, a number of recommendations for future are drawn.
These are categorised in groups and cited below:

Flow of information on technological advances (including forthcoming reports) on
synthetic biology)
o
The synthetic biology leadership council (SBLC) 55 looks on all aspects of
synthetic biology. Their views on food and feed synthetic biology products
and applications would be of interest.
o
Synthetic biology working group56 – The European Commission has
requested the Scientific Committee on Emerging and Newly Identified Health
Risks (SCENIHR), in association with the Scientific Committee on Consumer
Safety (SCCS) and with the Scientific Committee on Health and
Environmental Risks (SCHER) to deliver an opinion on Synthetic Biology
(Synthetic biology). This WG is currently deliberating the potential
applications of synthetic biology in various sectors and their implications for
regulatory risk assessment.
o
EU/US task force on biotechnology research57: The Synthetic Biology working
group was established in 2010 to foster exchange of views and collaboration
on scientific and technical progress in implementing synthetic biology
principles in such areas as standards, orthogonality, minimal genomes, ethics,
biosafety (including environmental safety), biosecurity, and education.
Following the background and stated needs the group already organized a
successful workshop58: Over the 2011-2015 period the working group will
focus on standardisation needs not met or realized yet via current practice; it
will follow ethical, legal and social issues in relation to the scientific and
technical progress of synthetic biology; and will pay special attention to the
contribution of synthetic biology to the different domains of biotechnology.
55
For more information on this see: https://connect.innovateuk.org/web/synthetic-biology-special-interestgroup/sblc-members
56
http://ec.europa.eu/health/scientific_committees/docs/synthetic_biology_mandate_en.pdf
57
http://ec.europa.eu/research/biotechnology/eu-us-task-force/index_en.cfm?pg=workinggroup#current
58
Standards in Synthetic Biology, Segovia, Spain, 2010
35 | P a g e
Calls encouraging coordinated research projects on synthetic biology
standardization of biological parts have recently been launched.
o
Keeping up to date with current technological development and available
literature, including EU FP6 and FP7 Synthetic Biology projects59: this could
be a challenge on its own as this field is progressing very quickly in many
different directions that could reflect variable food and feed products and
applications. Setting a core database or a data sharing platform to include all
available up to date published information could be an important step ahead.
Several research groups as well as regulatory agencies could have access to
such data sharing points; however, a number of key issues would need to be
addressed: cost of setting up such databases and data sharing platforms,
maintenance frameworks, evaluation and quality control of potential entries,
updating, organising and monitoring services via such databases and data
sharing points, etc.

Time considerations for ensuring that “fit-for-purpose” regulations are in place in
advance of applications for approval
o
Regulations take a long time to be shaped and agreed; thus one needs to start
preparing far in advance for case studies of “true” synthetic biology in the
food and feed sectors. At this point in time envisaging the full details of such
“true” synthetic biology products and applications as well as the pathway of
59
EU FP6 projects: SYNBIOLOGY: A European perspective on synthetic biology; BIOMODULARH2: Energy
project promises a new biotechnology; TESSY: foundations for a European synthetic biology; SYNPLEXITY:
Dynamics and complexity in synthetic protein networks (MOBILITY); CELLCOMPUT: – Biological computation
built on cell communication systems (NEST); SYNBIOSAFE: Safety and Ethical Aspects of Synthetic Biology.
EU FP7 projects: KBBE-2007-3-3-01 Synthetic Biology for the Environment (CSA-CA): Targeting
environmental pollution with engineered microbial systems a la carte (TARPOL); KBBE-2009-3-6-05: Synthetic
biology for biotechnological applications (CP-FP): Bacterial Synthetic Minimal Genomes for Biotechnology
(BASYNTHEC); KBBE.2011.3.6-03: Towards standardisation in Synthetic Biology (CP-IP): Standardization and
orthogonalization of the gene expression flow for robust engineering of NTN (new-to nature)
biological properties (ST-FLOW); KBBE.2011.3.6-04: Applying Synthetic Biology principles towards the cell
factory notion in biotechnology (CP-FP): Products from methanol by synthetic cell factories (PROMYSE) and
Code-engineered new-to-nature microbial cell factories for novel and safety enhanced bioproduction
(METACODE); KBBE.2011.3.6-06: Synthetic biology – ERA-NET. Call FP7-ERANET-2011-RTD: Development and
Coordination of Synthetic Biology in the European Research Area (ERASynBio); SiS-2008-1.1.2.1: Ethics and new
and emerging fields of science and technology: SYNTHETICS and SYBHEL; SiS.2012.1.2-1. Mobilisation and
Mutual Learning Action Plans; Acronym: SYN-ENERGY
36 | P a g e
the evolution of such products is very difficult. A possible way forward
would be to hold regular horizon scanning exercises on this subject.
o
Preparations for detailed risk assessments that are most probably burdened
with many uncertainties should be in place far in advance of applications for
approval of “true” synthetic biology products60. Such preparations should
involve the setup of assessed (national and international) experts’ networks
and structured methods to elicit qualitative and quantitative information.

Consideration of complexity of global food and feed trade for relevant synthetic
biology products and applications
o
Indirect effects on food and feed via other sectors, e.g. industrial applications
that produce waste that could be introduced into the food chain; to anticipate
potential effects one would need to have an updated, comprehensive view of
the whole system (i.e. describing the inputs and outputs of food/feed sectors).
This may involve the construction (and the regular updating) of conceptual
models to represent such systems; such exercises could form part of the
regular horizon scanning exercises mentioned above.

Clarity of introducing social and ethical inputs to regulatory processes and
stakeholder involvement
o
Ideally, synthetic biology aims for more sophisticated, efficient, sustainable
products and applications, so it would be imperative to ensure that
regulatory frameworks have the highest degree of clarity around how social
and ethical aspects will be taken into account as part of the approval process.
o
Comprehensive stakeholders’ involvement as early as possible would be
imperative to ensure that their knowledge, experience and views are taken
into account through the development of research, innovation and regulation
and to facilitate the development of “fit-for-purpose” regulatory frameworks.
A step towards the latter point would be to organise a day workshop with
invitees from NGOs and industry to deliberate about these issues, using the
findings of this project as one of the inputs.
60
For example, specific requirements about being explicit about uncertainties, and if possibly characterise and
quantify the uncertainties, should be included in the risk assessment.
37 | P a g e
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7. Annexes
Annex I-A: Deliverable 1 - Synopsis of current and projected products/applications
of synthetic biology for food/feed in the UK.
Synopsis of current and projected products/applications of synthetic
biology for food/feed in the UK
61
Contents
Item
1. Context
2. Criteria determining which literature items to include in Deliverable 1
3. Methodologies of literature search
4. Selected articles for potential Synthetic Biology food and feed products
and applications
5. Next steps – planning Deliverable 2
6. Annex I
Page
2
6
10
17
34
41
Image from: http://scitechdaily.com/synthetic-biology-circuits-perform-logic-functions-and-remember-theresults/
61
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1. Context
Synthetic Biology is a term that is broadly used to describe (a) the design and engineering of
biologically based materials as well as novel devices or whole systems, and (b) the re-design
of existing biological systems. It is an emerging bio-based technology which is quickly
establishing itself as a key enabling technology alongside nanotechnology (STOA workshop
report, 2012). Synthetic Biology has opened up a vast array of possibilities for applying an
engineering approach to biology, in which known biological entities can be (re)built to
perform a different or a new function, or completely new living organism can be synthesised
from scratch. The possibility of chemically synthesising biological entities, or a living
organism for a ‘designer’ function, is leading to potential precise engineering of metabolic
pathways, such as development of highly efficient therapeutic treatments (Khalil & Collins,
2010), and designing microorganisms for a specific or a novel function – such as clean-up of
the polluted environment (Schmidt, 2010). It is now anticipated, that Synthetic Biology
products/applications could provide elegant solutions to address a number of challenging
societal problems in the coming decades, such as food security (i.e. provide adequate
healthy and nutritious food to meet demand); ensuring accessibility to cheaper and highly
efficient pharmaceuticals; enhancement of production of “green” energy (e.g. biofuel
production); development of targeted therapies for chronic diseases, e.g. cancer and better
management of (food) waste (TNS-BMRB, 2010 - WP2; Flari & Chaudhry, 2012; STOA
workshop report, 2012; Hoskisson, 2012).
Potential Synthetic Biology products/applications in the food sector span a number of subsectors (OECD, 2009; European Commission, 2010a), in particular: (a) metabolites and health
products (e.g. vitamins) and processing aids in the manufacture of food and food
derivatives, such as nutraceuticals, probiotics and glycol nutrients used to raise the value of
certain foods or nutrient-enriched plants; (b) preservatives (e.g. Nisin); (c) flavours and
fragrances; (d) biosensors (e.g. artificial nose); (e) food waste processing. Food applications
enabled by new technologies, however, could carry new risks to health and the
environment; even if they do not, they could evoke large public sensitivities, and any new
application perceived to be risky is likely to raise concerns over consumer and environment
safety.
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During a recent expert workshop held at Fera on “New prospects and potential pitfalls of
Synthetic Biology” in September 2011 (Flari & Chaudhry, 2012), the participants noted that
possible adverse effects associated with Synthetic Biology products/applications could be
due to either intentional (i.e. targeted) or unintentional release. Additionally, the experts
distinguished real risks (e.g. recombination events; pathogen resistance) from perceived
risks, and acknowledged that, at this point in time, the uncertainties involved are too
many/too large to allow for a meaningful risk assessment of group/s of
products/applications. As a result, Synthetic Biology products/applications would need to be
assessed on a “case by case basis".
The fact that developments in emergent sciences and technologies often suffer setbacks in
early stages was also discussed during the workshop. Examples of such setbacks include the
moratorium on Genetically Modified Organisms in Europe (Amman, 2010; European
Commission 2010b); the stigmatisation of certain technologies in USA and Europe (e.g.
irradiation of food: Mehta, 2003); and nuclear technologies (Sjoberg, 2003; Vastchenko, 2006).
Furthermore, like other emergent technologies, the possibilities opened up by Synthetic
Biology have already raised certain ethical, safety and regulatory issues, in particular over
the potential risk of inadvertent or deliberate harmful use of the technology, e.g. in
developing biological weapons (IRGC, 2010). Experts agreed that all stakeholders interested
in Synthetic Biology products/applications should be involved in the development of
responsible regulatory frameworks, aiming towards shared, and if possible international
standards, and that any potential risks should be considered at the initial point of
development. During the workshop experts discussed the prerequisites to ensuring
responsible innovation. Although it was recognised that existing legislation for Genetically
Modified (GM) products/applications could be appropriate to regulate Synthetic Biology
products/applications, the experts expressed concerns that (a) following the GM
legislation/regulation example could block innovation, and (b) the regulations/legislation
currently in place may not suffice to address forthcoming product/application development
in the Synthetic Biology field.
A recent report from the European Group on Ethics in Science and New technologies
indicated that the legislative framework that might apply to Synthetic Biology is strictly
dependent on the potential applications of the products of this scientific sector (European
46 | P a g e
Group on Ethics in Science and New Technologies, 2009); therefore, it is expected to include
legal and policy provisions at different levels: (a) European Union (EU) legislation on
GMOs, biomedicine, bio-safety, chemicals, data protection and patents; (b) Global
provisions issued by the World Trade Organisation (WTO) and bio-safety standards issued
by the World Health Organisation (WHO); (c) International framework on ethics and human
rights. The same report indicated that the application areas of Synthetic Biology are already
regulated at EU level and Synthetic Biology products will have to comply with the existing
regulations.
For Synthetic Biology food/feed products/applications additional regulatory frameworks
may need to be taken into account, for example: (a) the Novel Foods Regulation (Regulation
(EC) No 258/97), including possibly the proposals for its replacement62; (b) Food Additives
legislation (i.e. Council Directive 89/107/EEC of 21 December 1988 on the approximation of
the laws of the Member States concerning food additives authorized for use in foodstuffs
intended for human consumption); (c) Flavourings legislation (Council Directive 88/388/EEC
of 22 June 1988 on the approximation of the laws of the Member States relating to
flavourings for use in foodstuffs and to source materials for their production). For such
products, the question becomes whether the current legislation and regulatory frameworks
are indeed appropriate and flexible enough to cover all of the anticipated needs.
Currently, Synthetic Biology research is being developed under laboratory conditions; in the
UK this research is regulated by the Health and Safety Executive. However, Synthetic
Biology products/applications may be ready for use outside the laboratory in the coming
years; depending on the applications and anticipated use, different regulatory bodies would
need to come on board, including FSA. The UK was amongst the first countries to recognize
the importance and potential impact by setting foundations on coordinating R&D and
engaging the public. As the pace of R&D in the Synthetic Biology field is expected to
increase very rapidly, and in order for the UK to keep on playing a distinguished leading
role in this field, it needs to consider pro-actively parallel development of (a) appropriate
regulatory schemes and (b) feasible implementation mechanisms. Such mechanisms must
62
Information available at: http://ec.europa.eu/food/food/biotechnology/novelfood/initiatives_en.htm
47 | P a g e
protect human health and the environment but must not be so burdensome as to
unnecessarily hinder responsible scientific development.
The project team developed a step-wise methodology to critically search the publicly
available information with the view to identifying literature items that would potentially
refer to current and imminent Synthetic Biology products and applications in the food and
feed sectors. Thereafter, the project team critically screened the identified literature items
and selected the ones that would be considered in depth, i.e. reviewed in depth and
categorised, in the next phase of the project.
48 | P a g e
2. Criteria determining what to include or exclude as information sources in
the synopsis
It is widely recognised that GM and Synthetic Biology are based on the same technologies
(e.g. genetic modification, genetic engineering), therefore one expects that at this point in
time, where the potential of Synthetic Biology just started to be unleashed, a large area of
potential overlap exists between GM and SynBio products and applications. However, at
this time, it is very difficult to know exactly how large this overlap between these two
science fields is, or where exactly does GM stop and Synthetic Biology start.
To address the objectives of the project it was considered crucial to try to set the boundaries
between GM products/applications and SynBio products/applications as clear as possible. In
doing so, we reviewed existent definitions of Synthetic Biology (shown in Table 1 below),
and developed a working understanding of what would constitutes a Synthetic Biology
product in food/feed as follows: “A Synthetic Biology product/application would include
substantially large63,64,65 synthetic parts of genetic material caused to function in a biological
system”. It is recognised that this phrase could be defined much more precisely (e.g. what is
considered as large?); nevertheless, such working understanding proved to be very valuable
as the project team was able to apply it as a “sieve” to filter out potentially interesting items
(see sections 3 and 4 of the report below).
Additionally, it is anticipated that a clear and concise definition of Synthetic Biology
products/applications for our project will enable us to define the boundaries between what
is perceived as a GM or a Synthetic Biology product/application. Further, this clarification
will also allow us to agree appropriate (non-zero) thresholds for the accidental or
adventitious presence of Synthetic Biology products in “conventional” products and to
determine suitable units of measurement.
If number of genes altered is taken into account: GM example in Mohamadzadeh et al., 2011: Regulation of
induced colonic inflammation by Lactobacillus acidophilus deficient in lipoteichoic acid. PNAS March 15, 2011 vol.
108 no. Supplement 1 4623-463012,13. Bacteria (i.e. Lactobacillus acidophilus) were genetically engineered not to
produce lipoteichoic acid. One gene was altered.
64[Online] Available at: http://www.pnas.org/content/108/suppl.1/4623.full.pdf+html?sid=9b45c08d-4ed7-41d9aebc-77f7f7beb8a1 [Last accessed 25 06 2013]
65 Media article in http://www.technologyreview.com/news/422571/genetically-engineered-probiotics/ [Last
accessed 25 06 2013]
63
49 | P a g e
Table 1: Collection of existent definitions for Synthetic Biology. Definitions are listed
chronologically.
Source
Definition
Key words / Focus
European
Commission
Report of a NEST
High-Level Expert
Group: “Synthetic
Biology Applying
Engineering to
Biology”
Synthetic biology is the engineering of
biology: the synthesis of complex,
biologically based (or inspired)
systems which display functions that
do not exist in nature. This
engineering perspective may be
applied at all levels of the hierarchy of
biological structures from individual
molecules to whole cells, tissues and
organisms. In essence, synthetic
biology will enable the design of
biological systems in a rational and
systematic way.
Engineering
Synthetic biology is the engineering of
biological components and systems
that do not exist in nature and the reengineering of existing biological
elements; it is determined on the
intentional design of artificial
biological systems, rather than on the
understanding of natural biology.
Engineering of biological
components and systems that do
not exist in nature
Synthetic biology is a new and rapidly
emerging discipline that aims at the
(re-)design and construction of (new)
biological systems.
(Re-)design of (new) biological
systems
Synthetic Biology is a new approach to
engineering biology, with an emphasis
on technologies to write DNA. Recent
advances make the de novo chemical
synthesis of long DNA polymers
routine and precise. Foundational
work, including the standardization of
DNA-encoded parts and devices,
enables them to be combined to create
programs to control cells. With the
development of this technology, there
is a concurrent effort to address legal,
social and ethical issues.
Technologies to write DNA
2005
Synthetic Biology
project EU FP666
2006
Synthetic Biology
3.067
June 2007
Synthetic Biology
4.068 (October 2008)
2008
Synthesis of complex,
biologically based systems that
display functions that not exist in
nature
Synthesis of complex,
biologically inspired systems
that display functions that do not
exist in nature
Re-engineering of existing
biological elements
Construction of (new) biological
systems
De novo synthesis of long DNA
polymers
Creates programs to control cells
[Online] Available at: http://www2.spi.pt/synbiology/documents/news/D11%20-%20Final%20Report.pdf [Last
accessed 24 06 2013]
67 [Online] Available at: http://www.syntheticbiology3.ethz.ch/index.htm [Last accessed 24 06 2013]
68 [Online] Available at: http://sb4.biobricks.org/field/ [Last accessed 24 06 2013]
66
50 | P a g e
Source
Definition
Key words / Focus
UK parliamentary
office for Science
and Technology
Post Note69
Synthetic biology aims to design and
build new biological parts and
systems or to modify existing ones to
carry out novel tasks.
Design and build of new
biological parts and systems
Synthetic Biology aims at designing
biological systems that do not exist in
nature using engineering principles or
re-designing existing ones to better
understand life processes, to generate
and assemble functional modular
components, and to develop novel
applications or processes.
Design biological systems that
do not exist
A definition of synthetic biology
should therefore include:
Identification, use and design of
biological parts totally or
partially artificial.
2008
Towards a
European Strategy
for Synthetic
Biology
EU FP670
Ethic report 71
1.The design of minimal
cells/organisms (including minimal
genomes);
Modifies existing biological parts
and systems to carry out novel
tasks.
Re-design existing ones
Generate and assemble
functional modular components
Develop novel applications and
processes
2. The identification and use of
biological ‘parts’ (toolkit);
3. The construction of totally or
partially artificial biological systems.
Synthetic Biology
org72
Synthetic Biology is (a) the design and
construction of new biological parts,
devices, and systems, and (b) the
redesign of existing, natural biological
systems for useful purposes.
Design of new biological parts,
devices and systems
Construction of new biological
parts, devices and systems
Redesign of existing, natural
biological systems
Prof Richard
Kitney for
“Synthetic Biology
From Science to
Governance:
A workshop
organised by the
European
Commission’s
Two complementary definitions for
SynBio: (a) designing and making
biological parts and systems that do
not exist in the natural world using
engineering principles, and (b)
redesigning existing biological
systems, again using engineering
principles.
Designing and making biological
parts and systems that do not
exist in the natural world using
engineering principles
Redesigning existing biological
systems using engineering
principles
Directorate[Online] Available at: http://www.parliament.uk/documents/post/postpn298.pdf [Last accessed 24 06 2013]
http://www.tessy-europe.eu/public_docs/TESSY-Final-Report_D5-3.pdf
71 [Online] Available at: http://ec.europa.eu/bepa/european-group-ethics/docs/opinion25_en.pdf [Last accessed 03
07 2013]
72 [Online] Available at: http://syntheticbiology.org/ [Last accessed 24 06 2013]
69
70
51 | P a g e
Source
Definition
Key words / Focus
Synthetic biology is the name given to
an emerging field of research that
combines elements of biology,
engineering, genetics, chemistry, and
computer science. The diverse but
related endeavors that fall under its
umbrella rely on chemically
synthesized DNA, along with
standardized and automatable
processes, to create new biochemical
systems or organisms with novel or
enhanced characteristics.
Combines elements of biology,
engineering, genetics, chemistry
and computer science.
Synthetic biology is the design and
engineering of biologically based
parts, novel devices and systems as
well as the redesign of existing,
natural biological systems.
Design of biologically based
parts, novel devices and systems
General for Health
& Consumers”73.
March 2010
Presidential
Commission for
the Study of
Bioethical Issues,
Report on
Synthetic
Biology74
2011
A synthetic
biology roadmap
for the UK75
2012
Relies on chemically synthesized
DNA
Relies on standardized and
automatable processes
Creates new biochemical
systems
Creates new organisms with
novel or enhanced characteristics
Engineering of biologically based
parts, novel devices and systems
Redesign of existing, natural
biological systems
UNICRI76
2012
Working
understanding of
Synthetic Biology
for Fera/FSA
Synthetic Biology is the deliberate
design of biological systems and living
organisms using engineering
principles
Design of biological systems
Short title: Extreme GM
Substantially long parts of
genetic material
Long title: Substantially large77,78,79
synthetic parts of genetic material
caused to function in a biological
Synthetic parts
[Online] Available at: http://ec.europa.eu/health/dialogue_collaboration/docs/synbio_workshop_report_en.pdf
[Last accessed 24 06 2013]
74 [Online] Available at: http://bioethics.gov/sites/default/files/PCSBI-Synthetic-Biology-Report-12.16.10_0.pdf
[Last accessed 24 06 2013]
75 [Online] Available at: http://www.rcuk.ac.uk/documents/publications/SyntheticBiologyRoadmap.pdf [Last
accessed 24 06 2013]
76 [Online] Available at:
http://www.unicri.it/in_focus/files/UNICRI%202012%20Security%20Implications%20of%20Synthetic%20Biology
%20and%20Nanobiotechnology%20Final%20Public-1.pdf [Last accessed 03 07 2013]
77 If number of genes altered is taken into account: GM example in Mohamadzadeh et al., 2011: Regulation of
induced colonic inflammation by Lactobacillus acidophilus deficient in lipoteichoic acid. PNAS March 15, 2011 vol.
108 no. Supplement 1 4623-463012,13. Bacteria (i.e. Lactobacillus acidophilus) were genetically engineered not to
produce lipoteichoic acid. One gene was altered.
78[Online] Available at: http://www.pnas.org/content/108/suppl.1/4623.full.pdf+html?sid=9b45c08d-4ed7-41d9aebc-77f7f7beb8a1 [Last accessed 25 06 2013]
79 Media article [Online] Available at: http://www.technologyreview.com/news/422571/genetically-engineeredprobiotics/ [Last accessed 25 06 2013]
73
52 | P a g e
Source
Definition
Key words / Focus
project 2013/2014
system.
Function as a biological system
53 | P a g e
3. Methodologies of literature search
A number of commercial databases covering food science, toxicology, agriculture and the
biosciences were searched. The various hosts by which Fera staff access the above databases
permit the creation of saved search logics which subsequently run automatically, thereby
providing alerting services. In addition to peer reviewed and commercial databases, we
searched “grey literature” sources, e.g. monitoring well established web information from
MIT synthetic biology working group in their synthetic biology community page 80,
Synthetic Biology project of Woodrow Wilson International Center for Scholars81, European
Union projects on Synthetic Biology82, J Craig Venter institute83, market research reports, and
media reports, e.g. Meltwater® media monitoring service which allows global monitoring of
the trade press and popular media. It is possible that web searching will identify relevant
market research reports84, although it may be not possible to acquire these within this
project’s life time as they could be very expensive. An initial list of information sources is
listed in Table 2 below.
References either initially or ultimately determined to be relevant and eligible for this review
(see next section) will, where possible, were entered into an Endnote or were produced as a
text report.
It was recognised from the beginning that developing a literature search strategy for the
purpose of this project would be a highly iterative process, and it was expected that this
would be further refined as the project progressed. Such searching is a compromise between
‘recall’ (getting everything) and ‘precision’ (getting just what you want) with the two being
inversely related85.
http://syntheticbiology.org/; http://biobricks.org/about-foundation/board-of-directors/
http://www.synbioproject.org/about/
82A list of EU FP6 and FP7 projects can be found in European Commission request for a joint scientific opinion on
Synthetic Biology [Online] Available at:
http://ec.europa.eu/health/scientific_committees/docs/synthetic_biology_mandate_en.pdf [Last accessed on 16 12
2013]
83 http://www.jcvi.org/cms/home/
84 SYNTHETIC BIOLOGY: LIVELIHOODS AND BIODIVERSITY VANILLA – extracted pages shown in Annex I.
85 http://www.creighton.edu/fileadmin/user/HSL/docs/ref/Searching_-_Recall_Precision.pdf
with the two being inversely related
80
81
54 | P a g e
Table 2: Sources of information searched in this project.
General science
databases
Patents
databases
Web of Knowledge Host
ISI Web of Science
BIOSIS,
CAB Abstracts,
Food Science and
Technology Abstracts
Zoological Record
ProQuest Dialog Host
AGRICOLA (AGRICultural
OnLine Access) 70-2013/Aug
CSA Life Sciences Abstracts
1966-2013/Jul
ELSEVIER BIOBASE 19942013/Aug
EMBASE 1993-2013/Aug 15
Pascal - French National
Research Council 19732013/Aug
Other
Derwent Innovations
Index – the is the
equivalent of Derwent
World Patent index as
specified in the original
proposal
(NB: many of the other
databases listed here
include patent records)
Grey Literature
FEDRIP - Federal Research In
Progress - 2013/Jun
NTIS (US National Technical
Information Service) 19642013/Aug
Google advanced search of the
web.
Other
Meltwater®
media monitoring
service
ReportLinker –
Market Research
reports
3.1
Search strategy – draft 1
The initial literature search strategy was planned to comprise three major steps designed to
search titles and subject headings in a number of databases (databases searched are shown
in Table 2 below).
.Combinations of key words were used, starting with a broad screening on terminology
related to synthetic biology (step 1) and then narrowing down to products and applications
in the food and feed areas (step 2; terms taken into account in this step are shown in Table 2
55 | P a g e
below). Finally, the results would be focused to the working understanding of synthetic
biology for this Fera/FSA project (step 3). This preliminarily version of the protocol for the
literature search (i.e. strategy steps and proposed search strings) is shown graphically in
Figure 1.
Figure 1: Graphical representation of the initial search strategy planned for the project.
3.2
Search strategy – draft 2
Example literature searches followed the strategy outlined above; the results obtained
indicated that the particular key search strings were too wide-ranging. For example when
only a very small number of databases were searched (n=3), step 1 produced 3,462,763
documents whereas a combination of step 1 and step 2 produced 14,424,891 documents.
When step 1 results were combined with one search string from step 2 (i.e. food) the search
produced 166,315 documents. However, when only the terms “synbio*” or “synthetic
biology” were combined with the term “food” (search string from step 2) the number of
documents found were reduced to 961. This amended strategy initially looked attractive as it
reduced the number of documents to a far more pragmatic level; nevertheless, it did raise
the question of whether this strategy may have missed products and applications that were
56 | P a g e
not trademarked as “synthetic biology” ones by the developers and authors. Additionally, it
did not rule out GM products and applications trademarked as “synthetic biology”.
We decided to try an alternative approach and identify further key words and search strings
based on: (a) published work of identified synthetic biology scientific hubs, for example:
Haseloff lab in University of Cambridge86; Institute of Biological Engineering87; Pamela
Silver laboratory in Harvard University88; Keith Waldron in Institute of Food Research89,90;
Synthetic Biology Institute in University of California, Berkeley91; the Centre for Synthetic
Biology and Innovation (CSynBI) at Imperial College London92; Syntegron project website93
(6 laboratories involved94); UNICRI 2012 report95; Registry of standard biological parts96;
Synthetic Biology based on standard parts97; forthcoming conference of industry
(engineering; investors; consulting; liaisons between academia and industry)98; (b) a number
of key Synthetic Biology documents identified by the project team members 99.
It was decided that information sources, apart from patents, would be screened back to 2003,
as it was thought that any papers dated earlier than this year would most probably refer to
GM rather than Synthetic Biology. The project team decided not to restrict the search for
relevant patents chronologically.
http://data.plantsci.cam.ac.uk/Haseloff/
http://www.ibe.org/engineering-for-life/body-of-knowledge.html
88 http://www.openwetware.org/wiki/Silver_Lab#Research_Information
89 http://www.ifr.ac.uk/profile/keith-waldron.asp
90 http://www.foodnavigator.com/Science-Nutrition/International-research-project-to-develop-synthetic-yeastfor-industry-applications
91 http://synbio.berkeley.edu/
92 http://www3.imperial.ac.uk/syntheticbiology
93 http://www.syntegron.org/
94 University of Glasgow - Rosser Lab: Dr. Susan Rosser; Dr. Sean Colloms; Northwestern University - Leonard
Lab: Dr. Josh Leonard; Mr. Andrew Scarpelli; University of California, Berkeley - Keasling Lab: Prof. Jay
Keasling; Dr. Josh Gilmore; John Innes Centre - Osbourn Lab; Prof. Anne Osbourn; Dr. Nan Yu; University of
Exeter - Bates Lab: Prof. Declan Bates; Dr. Adam Spargo; Imperial College London - Freemont Lab: Prof. Paul
Freemont; Dr. James MacDonald; Dr. Sarah Butcher; Dr. James Abbott
95 UNICRI, 2012: Security implications of synthetic biology and nanobiotechnology – a risk and response
assessment of advances in biotechnology. [Online] Available at:
http://www.unicri.it/in_focus/files/UNICRI%202012%20Security%20Implications%20of%20Synthetic%20Biology
%20and%20Nanobiotechnology%20Final%20Public-1.pdf [Last accessed 16 12 2013]
96 http://parts.igem.org/Help:Standards
97 http://igem.org/About
98 http://synbiobeta.com/sf2013/
99 These documents will be considered for the objectives of Deliverables 2, 3 and 4.
86
87
57 | P a g e
Draft 2 of our search strategy comprised only the first two steps outlined in Figure 1 as its
aim was to refine as much as possibe the terms in these steps. The search strings we
employed are shown in Annex I.
We screened all available databases (as shown in Table 2 below) employing the above
selected terms; our search produced 2643 artciles and 92 patents as potentially relevant and
interesting items. Thereafter we critical evaluated the results published in years 2013, 2012,
and 2011; our evaluation indicated that although we identified a number of potentially
relevant articles the great majority of the outputs were considered as non-relevant.
3.3
Search strategy – final
Results following the application of the 2nd draft of the search strategy indicated that we
need to re-evaluate the search strings, mainly in order to reduce the number of non-relevant
articles. We introduced the desirable search strings in three concepts, i.e. Synthetic Biology,
Application and Food/Feed sectors, and included a fourth concept that referred to terms we
did not desire to see in the identified items (Table 3). The concept of Synthetic Biology was
heavily edited (compared with the search strings included in step 1 in drafts 1 and 2 of the
search strategy) in order to exclude terms that were very generic.
Table 3: The final search strategy is illustrated in the table below. The search was unfolded
in four concepts: concept one took into account refined synthetic biology affiliated terms;
concept two took into account the types of applications that are of interest for the project;
concept three focused the results to food and feed sectors; concept four refined the results by
introducing “exclusion” terms in order to decrease the number of non-relevant items in the
search outputs.
Concepts
1: Synthetic Biology
2: Applications
3: Food / Feed
4: Exclude terms
Allylix
Additive
Crop
Biosensor*
Biobrick
Antioxidant
Feed
Environment*
Bioengineer or bioengineering
Colour
Feedstock
Impact*
58 | P a g e
Concepts
1: Synthetic Biology
2: Applications
3: Food / Feed
4: Exclude terms
Biological pathway synthesis
Emulsifier
Food
Medical
Biotransformation
Enzyme
Nanotechnology
Bottom-up
Fermentation
Nanotechnologies
Cell function reprogram or
product
Pharmaceutical
reprogramming
Flavour
Waste
Cellular chassis
Flavour enhancer
Chemical synthetic biology
Fragrance
Designer strains
Gelling agent
Directed evolution
Health food
DNA assembly part
Ingredient
DNA based device
Nutraceutical
construction
Metabolite
DNA modular construction
Preservative
Engineering a living system
Probiotic
Engineering cells
Processing aid
Evolutionary engineering
Stabiliser
Genetic brick
Sweetener
Genetic circuits
Thickener
GenoCAD
Genome driven cell
engineering
Genome engineering
Genome scale DNA synthesis
Industrial microorganism
Metabolic engineering
Metabolite channelling
Microbial cell factory or
Microbial cell factories
Minimal genome
Model synthetic
Multi-enzyme one pot
Pathway engineering
59 | P a g e
Concepts
1: Synthetic Biology
2: Applications
3: Food / Feed
4: Exclude terms
Plasmid backbone
Protocell
Protocell creation
Recombinant gene cluster
Recombineering
Standardised genetic modular
part
Substrate channelling
Synthetic biology
Synthetic DNA
Synthetic enzyme
Synthetic gene circuits
Synthetic gene transcription
Synthetic genome
Synthetic genomics
Synthetic meat
Synthetic metabolic pathway
Top-down
60 | P a g e
4.
Selected articles for potential Synthetic Biology food and feed products and
applications
The final search strategy described above was employed to search available information (as
indicated in Table 2 above); 713 articles and 108 patents were identified as potentially
relevant to the objectives of the project. These were critically screened by the project team
members to identify the most relevant ones, and the working understanding for Synthetic
Biology (as defined above in page 6) was applied in order to screen out products and
applications that were typically GM. Some examples of such “rejected” products and
applications are shown below in Table 4.
Table 4: Examples of “rejected” products and applications as they were considered to
describe products and applications that were typically GM.
Article
GM technology
Cahoon, E. B. and K. M. Schmid
(2008). Metabolic Engineering of the
Content and Fatty Acid Composition
of Vegetable Oils. Bioengineering and
Molecular Biology of Plant Pathways.
H. J. Bohnert, H. Nguyen and N. G.
Lewis. 1: 161-200.
Carrari, F., R. Asis, et al. (2007). "The
metabolic shifts underlying tomato
fruit development." Plant
Biotechnology 24(1): 45-55.
Total oil production has been increased moderately by
overexpression of genes for the first and last steps of oil
synthesis, acetyl-CoA carboxylase (ACCase), and
diacylglycerol acyltransferase (DGAT), respectively.
Changfu, Z., S. Naqvi, et al. (2008).
"Combinatorial genetic transformation
generates a library of metabolic
phenotypes for the carotenoid
pathway in maize." Proceedings of the
National Academy of Sciences of the
United States of America 105(47):
18232-18237.
Delaney, B. (2007). "Strategies to
evaluate the safety of bioengineered
foods." International Journal of
Toxicology 26(5): 389-399.
A review is presented of recent reports focused on the
identification of key points on the metabolic regulation
underlying tomato fruit development. Additionally, an
overview of the combined application of metabolic and
transcriptional profiling, aimed at identifying candidate
genes for modifying metabolite contents, is discussed in
the context of the usefulness for tomato breeding
programmes.
Specifically, 5 carotenogenic genes controlled by different
endosperm-specific promoters were transferred into a
white maize (corn) variety deficient for endosperm
carotenoids synthesis.
A number of genetically modified (GM) crops
bioengineered to express agronomic traits including
herbicide resistance and insect tolerance have been
commercialized. Safety studies conducted for the whole
grains and food and feed fractions obtained from GM
crops (i. e., bioengineered foods) bear similarities to and
61 | P a g e
Article
Fujisawa, M. and N. Misawa (2010).
Enrichment of Carotenoids in
Flaxseed by Introducing a Bacterial
Phytoene Synthase Gene. Plant
Secondary Metabolism Engineering:
Methods and Applications. A. G.
FettNeto. 643: 201-211.
Mierau, I., P. Leij, et al. (2005).
"Industrial-scale production and
purification of a heterologous protein
in Lactococcus lactis using the nisincontrolled gene expression system
NICE: The case of lysostaphin."
Microbial Cell Factories 4.
Palazon, J., E. Moyano, et al. (2006).
Tropane alkaloids in plants and
genetic engineering of their
biosynthesis.
Wei, X.-X. and G.-Q. Chen (2008).
Applications of the VHb gene vgb for
improved microbial fermentation
processes. Globins and Other Nitric
Oxide-Reactive Proteins, Pt A. R. K.
Poole. 436: 273-+.
GM technology
distinctive differences from those applied to substances
intentionally added to foods (e. g., food ingredients).
A phytoene synthase gene (crtB) derived from a soil
bacterium Pantoea ananatis (formerly called Erwinia
uredovora) strain 20D3 was introduced into L.
usitatissimum WARD cultivar. The resulting transgenic
flax plants formed orange seeds, which contained
phytoene, alpha-carotene, beta-carotene, and lutein. The
Example that shows that nisin-regulated gene expression
in L. lactis can be used at industrial scale to produce large
amounts of a target protein, such as lysostaphin.
The pmt gene of Nicotiana tabacum under the control of
the CaMV 35S promoter was introduced into the genome
of two scopolamine-rich plant species, Datura metel and a
Duboisia hybrid, together with the T-DNA of the Ri
plasmid from Agrobacterium rhizogenes.
To alleviate the defects of hypoxic conditions, Vitreoscilla
hemoglobin (VHb) has been used to enhance respiration
and energy metabolism by promoting oxygen delivery.
The items that were considered as relevant to the project’s objectives are shown below in
Table 5. These include items that were considered as Synthetic Biology products and
applications (highlighted in green), items that refer to products and applications that are
based on methods which are rather closer to GM than Synthetic Biology, albeit not strictly
GM, and finally items that were considered as valuable because of the methodologies
described that would perhaps lead to the development of food / feed related Synthetic
Biology products and applications in the future.
62 | P a g e
Table 5: Inventory of articles identified as relevant to the aims and objectives of the study. The degree of relevance of each article is indicated
by the colour of the symbol on the left of each title. Thus:

Green colour indicates high relevance.

Blue colour indicates that the project team considered the methods followed in the article closer to GM than Synthetic Biology.

Red colour indicates that the article was considered as valuable because of the methodologies described that would perhaps lead to the
Aharoni, A. and E. Lewinsohn (2010). Genetic
engineering of fruit flavors. Handbook of fruit and
vegetable flavors: 101-114. ISBN 978-0-470-22721-3
Akhila, A. (2007). "Metabolic engineering of
biosynthetic pathways leading to isoprenoids: Monoand sesquiterpenes in plastids and cytosol." Journal
of Plant Interactions 2(4): 195-204.
Alchihab, M., J. Destain, et al. (2010). "Production
of aroma lactones by yeasts." Biotechnologie
Agronomie Societe Et Environnement 14(4): 681-691.
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Colour agent
Antioxidant
Additive
Article or patent
Crop or Crop derived product
development of food / feed related Synthetic Biology products and applications in the future.
x
x
x
x
63 | P a g e
Azadi, P., N. V. Otang, et al. (2010). "Metabolic
engineering of Lilium x formolongi using multiple
genes of the carotenoid biosynthesis pathway." Plant
Biotechnology Reports 4(4): 269-280.
Baeumchen, C., A. H. F. J. Roth, et al. (2007). "Dmannitol production by resting state whole cell
biotransformation of D-fructose by heterologous
mannitol and formate dehydrogenase gene
expression in Bacillus megaterium." Biotechnology
Journal 2(11): 1408-1416.
Barghini, P., D. Di Gioia, et al. (2007). "Vanillin
production using metabolically engineered
Escherichia coli under non-growing conditions."
Microbial Cell Factories 6. DOI 13
10.1186/1475-2859-6-13
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
x
x
x
64 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Baumgartner, F., L. Seitz, et al. (2013).
"Construction of Escherichia coli strains with
chromosomally integrated expression cassettes for
the synthesis of 2 '-fucosyllactose." Microbial Cell
Factories 12. DOI 40 10.1186/1475-2859-12-40
Blankschien, M. D., J. M. Clomburg, et al. (2010).
"Metabolic engineering of Escherichia coli for the
production of succinate from glycerol." Metabolic
Engineering 12(5): 409-419.
Bovy, A., E. Schijlen, et al. (2007). "Metabolic
engineering of flavonoids in tomato (Solanum
lycopersicum): the potential for metabolomics."
Metabolomics 3(3): 399-412.
Brochado, A. R., C. Matos, et al. (2010). "Improved
vanillin production in baker's yeast through in silico
design." Microbial Cell Factories 9. 84
DOI 10.1186/1475-2859-9-84
x
x
x
65 | P a g e
Burgess, C., W. Sybesma, et al. (2003).
"Characterisation and improvement of riboflavin
production in Lactococcus lactis." Abstracts of the
General Meeting of the American Society for
Microbiology 103: O-088.
Burja, A. M. and H. Radianingtyas (2005). "Marine
microbial-derived nutraceuticals biotechnology: an
update." Food Science & Technology 19(1): 14-16.
Cimini, D., K. R. Patil, et al. (2005). "Metabolic
engineering of succinic acid production in
Saccharomyces cerevisiae." Journal of Biotechnology
118: S118-S118.
Cunningham, F. (2003). "Metabolic engineering of
pathways leading to fat-soluble vitamins and
antioxidants in plants: What have we learned and
where are we going?" Hortscience 38(5): 715-715.
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
x
x
x
x
66 | P a g e
Damude, H. G. and A. J. Kinney (2007). Metabolic
engineering of seed oil biosynthetic pathways for
human health. Verpoorte, R., Alfermann, A. W.,
Johnson, T. S. Applications of Plant Metabolic
Engineering. 978-1-4020-6030-4(H). DOI 10.1007/9781-4020-6031-1_10
Dyer, J. M., D. C. Chapital, et al. (2003). "Metabolic
engineering of bakers' yeast for production of valueadded lipids." Abstracts of Papers American
Chemical Society 225(1-2): 115-AGFD 115.
Field, D., M. Begley, et al. (2012). "Bioengineered
Nisin A Derivatives with Enhanced Activity against
Both Gram Positive and Gram Negative Pathogens."
PLoS ONE 7(10).
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
x
x
x
67 | P a g e
x
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
x
Nutraceutical
x
Metabolite
Ingredient (incl. fermentation product)
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Flavouring agent
x
Health product
Gaspar, P., A. R. Neves, et al. (2008). Engineering
and re-routing of the metabolism of lactic acid
bacteria. Mayo, B., Lopez, P., Perez-Martinez, G.
Molecular aspects of lactic acid bacteria for
traditional and new applications. ISBN 265-289. 97881-308-0250-3
Han, X.-y., D.-q. Sun, et al. (2007). "Gene regulation
to lactic acid bacteria for increasing production of
flavor metabolite." Weishengwu Xuebao 47(6): 11051109.
Hendrawati, O., H. J. Woerdenbag, et al. (2010).
"Metabolic engineering strategies for the
optimization of medicinal and aromatic plants:
realities and expectations." Zeitschrift Fur Arznei- &
Gewurzpflanzen 15(3): 111-126.
Iijima, Y. (2010). Metabolic factory for flavors in
fruits and vegetables. Handbook of fruit and
vegetable flavors: 705-727. ISBN 978-0-470-22721-3
Antioxidant
Additive
Article or patent
x
x
68 | P a g e
Jin-Ho, C., R. Yeon-Woo, et al. (2005).
"Biotechnological production and applications of
coenzyme Q10." Applied Microbiology and
Biotechnology 68(1): 9-15.
Jung, W. s., I.-M. Chung, et al. (2003).
"Manipulating isoflavone levels in plants." Journal of
Plant Biotechnology 5(3): 149-155.
Kim, S.-H., S.-Y. Kim, et al. (2009). "Biosynthesis of
Polyunsaturated Fatty Acids: Metabolic Engineering
in Plants." Journal of Applied Biological Chemistry
52(3): 93-102.
Kong M.K., Lee P.C. (2011) “Metabolic engineering
of menaquinone-8 pathway of Escherichia coli as a
microbial platform for vitamin K production.”
Biotechnology and Bioengineering 108(8): 1997–2002
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
x
x
x
x
69 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Koopman, F., J. Beekwilder, et al. (2012). "De novo
production of the flavonoid naringenin in
engineered Saccharomyces cerevisiae." Microbial
Cell Factories 11. DOI 155
10.1186/1475-2859-11-155
Lee, J. H. and D. J. O'Sullivan (2008). "Metabolic
Engineering of Lactococcus lactis for the
Development of a One-Step Bioconversion of Lactose
into Tagatose." Abstracts of the General Meeting of
the American Society for Microbiology 108: 473-473.
Lei, Z., W. Zinan, et al. (2007). "Metabolic
engineering of plant L-ascorbic acid biosynthesis:
recent trends and applications." Critical Reviews in
Biotechnology 27(3): 173-182.
Lemuth, K., K. Steuer, et al. (2011). "Engineering of
a plasmid-free Escherichia coli strain for improved in
vivo biosynthesis of astaxanthin." Microbial Cell
Factories 10. DOI 29
10.1186/1475-2859-10-29
x
x
x
x
x
70 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Lewinsohn, E., R. Davidovich-Rikanati, et al.
(2010). Functional genomics for the discovery of
genes affecting lemon basil aroma and their use in
flavor engineering of tomato. ISHS Acta
Horticulturae 860: IV International Symposium on
Breeding Research on Medicinal and Aromatic
Plants - ISBMAP2009
x
Liu, N. and S. Zheng (2010). "Advances in
biological functions and biosynthesis regulation of
tocotrienols." Guangxi Zhiwu / Guihaia 30(1): 122159.
Lorence, A. and C. L. Nessler (2007). Pathway
engineering of the plant vitamin C metabolic
network. Verpoorte, R., Alfermann, A. W., Johnson,
T. S. Applications of Plant Metabolic Engineering:
1970217. ISBN 978-1-4020-6030-4(H)
x
x
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Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Lu, C.-H., J.-H. Choi, et al. (2011). "LaboratoryScale Production of Tomato Carotenoids Using
Bioengineered Escherichia coli." FASEB Journal 25.
x
Luo, H., J. Song, et al. (2013). "Cloning and
expression analysis of a key device of HMGR gene
involved in ginsenoside biosynthesis of Panax
ginseng via synthetic biology approach." Acta
Pharmaceutica Sinica 48(2): 219-227.
Mattoo, A. K., V. Shukla, et al. (2010). Genetic
Engineering to Enhance Crop-Based Phytonutrients
(Nutraceuticals) to Alleviate Diet-Related Diseases.
Bio-Farms for Nutraceuticals: Functional Food and
Safety Control by Biosensors. M. T. Giardi, G. Rea
and B. Berra. 698: 122-143.
x
x
72 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Mehrotra, S., L. U. Rahman, et al. (2010). "An
extensive case study of hairy-root cultures for
enhanced secondary-metabolite production through
metabolic-pathway engineering." Biotechnology and
Applied Biochemistry 56: 161-172.
x
Melzer, G., M. E. Esfandabadi, et al. (2009). "Flux
Design: In silico design of cell factories based on
correlation of pathway fluxes to desired properties."
Bmc Systems Biology 3: 120
Misawa, N. (2009). "Pathway engineering of plants
toward astaxanthin production." Plant
Biotechnology 26(1): 93-99.
x
Moldrup, M. E., F. Geu-Flores, et al. (2011).
"Modulation of sulfur metabolism enables efficient
glucosinolate engineering."BMC Biotechnology
11(12): (31 January 2011)-(2031 January 2011).
x
73 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Park, J. H. and S. Y. Lee (2010). "Metabolic
pathways and fermentative production of Laspartate family amino acids." Biotechnology Journal
5(6): 560-577.
Peiru, S., E. Rodriguez, et al. (2008). "Metabolically
engineered Escherichia coli for efficient production
of glycosylated natural products." Microbial
Biotechnology 1(6): 476-486.
Reuben, S., L. J. Cseke, et al. (2006). Molecular
biology of plant natural products. Cseke, L. J.,
Kirakosyan, A., Kaufman, P. B., Warber, S. L., Duke,
J. A., Brielmann, H. L. Natural products from plants:
165-202. ISBN 0-8493-2976-0\978-0-8493-2976-0
x
x
74 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Sato, F. and K. Matsui (2012). Engineering the
biosynthesis of low molecular weight metabolites for
quality traits (essential nutrients, health-promoting
phytochemicals, volatiles, and aroma compounds).
Altman, A, Hasegawa, P. M., Plant Biotechnology
and Agriculture: Prospects for the 21st Century, 443461. 978-0-12-381467-8.
x
x
x
So-Yeon, S., H. Nam Soo, et al. (2011). "Production
of resveratrol from p-coumaric acid in recombinant
Saccharomyces cerevisiae expressing 4coumarate:coenzyme A ligase and stilbene synthase
genes." Enzyme and Microbial Technology 48(1): 4853.
x
Sun-Young, K., C. Oksik, et al. (2012). "Artificial
biosynthesis of phenylpropanoic acids in a tyrosine
overproducing Escherichia coli strain." Microbial
Cell Factories 11(Dec.): 153-153.
x
75 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Thanasomboon, R., D. Waraho, et al. (2012).
Construction of synthetic Escherichia coli producing
s-linalool. Proceedings of the 3rd International
Conference on Computational Systems-Biology and
Bioinformatics. J. H. Chan, A. Meechai and C. K.
Kwoh. 11: 88-95.
Van der Straeten et al. (2010) Patent Application
Publication US 2010/0183750 A1: Fortification of
plants with folates by metabolic engineering
x
x
Wang, Y., H. Chen, et al. (2010). "Metabolic
engineering of resveratrol and other longevity
boosting compounds." Biofactors 36(5): 394-400.
Watts, K. T., P. C. Lee, et al. (2006). "Biosynthesis of
plant-specific stilbene polyketides in metabolically
engineered Escherichia coli." BMC Biotechnology
6(22): (21 March 2006)-(2021 March 2006).
x
76 | P a g e
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl. fermentation product)
Health product
Gelling agent
Fragrance (relevant to food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived product
Colour agent
Antioxidant
Additive
Article or patent
Won-Heong, L., P. Panchalee, et al. (2012). "Whole
cell biosynthesis of a functional oligosaccharide, 2′ fucosyllactose, using engineered Escherichia coli."
Microbial Cell Factories 11(April): 48-48.
x
Yu, O. (2009). "Genetic regulation and metabolic
engineering of isoflavone biosynthesis." Abstracts of
Papers of the American Chemical Society 238: 294294.
x
Yue, A.-Q. and R.-Z. Li (2009). "Metabolic
Engineering to Produce Novel Renewable Industrial
Fatty Acids in Oilseeds." Zhongguo Shengwu
Huaxue yu Fenzi Shengwu Xuebao 25(6): 501-509.
x
Zvi, M. M. B., B. Spitzer, et al. (2006). Navigating the
network of floral scent production. Acta
Horticulturae. A. Mercuri and T. Schiva: 143-154.
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5. Next steps – planning Deliverable 2
5.1
Identifying experts for elicitation of necessary information
These initial peer reviewed searches enabled us to identify relevant science hubs in this area;
it may be possible that we will need to communicate directly with principal investigators
involved in the development of products and applications to find out prospective timelines.
Additionally, web searching results indicate the following science hubs: California Institute
of Technology, USA; Gate Fuels Inc, USA; Graz University of Technology, AU; Harvard
university, USA; Swiss Federal Institute of Technology, CZ; University of California, USA;
University of Groningen, NL; United States Department of Agriculture (USDA), USA;
University of Illinois, USA; University of Illinois Urbana Champaign, USA; University of
London, UK. Further, we identified potential industry links (shown in Table 6 below).
In addition, we may need to elicit information (possibly via remote surveys) that would help
us identifying potential points in food and feed production and consumption chains where
synthetic biology can provide relatively inexpensive solutions within certain timeframes, i.e.
in the short term, within next five years, and in the medium term, between five and ten
years.
5.2
Review and classification of identified items in Deliverable 1
Following the completion of Deliverable 1 (i.e. synopsis of potential Synthetic Biology
food/feed products/applications) we would explore one or more potential schemes to
classify / group the identified Synthetic Biology food/feed products/applications. A number
of possible criteria will be taken into account, for example (a) technologies employed in the
development of such products, (b) the likelihood of each product/application to be marketed
within the next 5-10 years, (c) potential health risks, (d) potential health benefits, (e) social
perceptions, (f) route/mode of delivery and potential food matrices to be employed, etc.
It is anticipated that categorizing/grouping the potential Synthetic Biology food/feed
products/applications would enable the team to select representative case studies to
facilitate next steps of the research, in particular objective 3 (i.e. review of regulatory
frameworks currently in place in terms of “fitness for purpose” to regulate Synthetic Biology
food/feed products/applications).
78 | P a g e
The robustness and usefulness of the classification/grouping schemes will be discussed with
all project team members during an expert elicitation exercise. During the same exercise we
will identify the factors potentially influencing their development, and evaluate likelihood
of different groups of products/applications being commercialised in the near future, i.e. 510 years time.
We would assign the identified Synthetic Biology food/feed products/application to the
classification/grouping schemes, with the view to selecting representative case studies. The
final selection of case studies would be decided through discussions with Molecular Biology
and Computational Biology experts at Fera, and it is anticipated that it will depend on the
availability of detailed reliable information. Examples of potential case studies could include
flavourings, e.g. Vanillin100; food preservatives, e.g. Nisin101; artificial sweeteners.
It is envisaged that for each selected case study the project team would develop an
information sheet that would include details on (a) technologies involved in the
development of the product, (b) application/s and expected effects (c) potential health risks,
(d) potential benefits and information on alternatives, (e) current status of development, (f)
academic institutes and industries involved, and (h) relevant uncertainties that burden the
assessment of the product/application.
(a) Development of Yest Cell Factory for Vanillin Production by using Synthetic Biology and Metabolic
Engineering Tools (Online) Available at: http://orbit.dtu.dk/en/projects/development-of-yest-cell-factory-forvanillin-production-by-using-synthetic-biology-and-metabolic-engineering-tools%28e0692685-1761-4bea-bca5511e69644de0%29.html - Last accessed on 10/09/2012
(b) A Submission to the Convention on Biological Diversity’s Subsidiary Body on Scientific, Technical and
Technological Advice (SBSTTA) on the Potential Impacts of Synthetic Biology on the Conservation and
Sustainable Use of Biodiversity (Online) Available at: https://www.cbd.int/doc/emerging-issues/Int-Civil-SocWG-Synthetic-Biology-2011-013-en.pdf - Last accessed on 10/09/2012
101 Synthetic biology to obtain novel antibiotics and optimized production systems (SYNMOD) [Online] Available
at: www.esf.org/activities/eurocores/running-programmes/eurosynbio/projects-crps/synmod.html [Last accessed
on 10/09/2012]
100
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Table 6: Industry potential links for the development of Synthetic Biology products and application s102.
Company
Genome compiler
EvolveMol
Biodesic
Integrated DNA technologies
On point capital
Lawrence Berkeley National
Laboratory
Lux Capital
LS9
DNA 2.0
Transcriptic
Caribou Biosciences
Angel investor for start-up
businesses
Country
USA
Ginkgo BioWorks
USA
http://ginkgobioworks.com/
Sample6 Technologies
USA
http://sample6.com/
USA
http://www.flagshipventures.com/
USA
http://www.pronutria.com/
VentureLab /Flagship
Innovation Foundry
Wired magazine; Extreme Files
Blog
Gen9
USA
USA
USA
Company website
http://www.genomecompiler.com/
http://www.evolvemol.com/
http://www.biodesic.com/
http://www.idtdna.com/site
http://www.onpointcapital.com/
Type / Products
Engineering
Engineering
Consulting - Technical
Engineering
Consulting - Financial
USA
http://www.lbl.gov/
Government  Industry
USA
USA
USA
USA
USA
http://www.luxcapital.com/
http://www.ls9.com/
http://www.dna20.com/
https://www.transcriptic.com/
http://www.cariboubiosciences.com/
Investment company
Industrial biotechnology
Gene synthesis
Cloning
Biotechnology
USA
USA
USA
Links with Food / Feed products
Investor
http://www.wired.com/wiredscience/
theextremofiles/
http://gen9bio.com/
Engineering
(microorganisms)
Diagnostics
Institutional platform
for investment
Flag company:
Pronutria
Flavor and fragrance companies103
Synthetic-biology diagnostic system104
Therapeutic nutrients
Writer
Gene synthesis
http://synbiobeta.com/sf2013/
Ginkgo engineers deliver scale-up-ready organisms in six months for the production of renewable fine and specialty chemicals. Our customers include sugar refiners, flavor
and fragrance companies, and other producers of fine chemicals.
104 Sample6 is the world’sfirst synthetic-biology based bacteria diagnostic system capable of enrichment-free detection; see also Annex II for more information for products of
this company.
102
103
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Deputy Director of Practices
for the National Science
Foundation Synthetic Biology
Engineering Research Center
(SynBERC)
Breakout Labs
Angel investor for start-up
businesses
Blue Marble
Synthace
GenScript
Genomatica
105
USA
http://www.synberc.org/content/rese
arch
USA
http://www.breakoutlabs.org/
USA
USA
UK
USA
USA
Investor
Investor
http://bluemarblebio.com/
http://www.synthace.com/
http://www.genscript.com/
http://www.genomatica.com/
Food industry
Engineering
Engineering
Sustainable chemicals
Variable products105
http://bluemarblebio.com/products/
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6. Annex I: List of search strings employed at the 2nd draft of search strategy.
Draft 2 of our search strategy comprised only the first two steps outlined in
Figure 1.
STEP 1: “Allylix” OR “biobrick” OR “biobrick device” OR “biobrick part” OR “biobrick
standard” OR “biocatalysis” OR “bioengineer or bioengineering “ OR “biological pathway
synthesis “ OR “biosensor” OR “biotransformation” OR “bottom-up” OR “branched chain
amino acid” OR “cell autonomous circuits” OR “cell free synthetic pathway” OR “cell
factory or cell factories” OR “Cell function reprogram or reprogramming “ OR “cell
membrane” OR “coimmobilization” OR “cellular chassis “ OR “chemical synthetic biology”
OR “designer strains” OR “directed evolution” OR “DNA assembly part “ OR “DNA-based
device construction “ OR “DNA modular construction “ OR “DNA synthesis” OR
“engineering cells “ OR “engineered E.coli strain” OR “engineering a living system “ OR
“enzyme complex” OR “evolutionary engineering” OR “feedback inhibition” OR “fed batch
fermentation” OR “genocad” OR “genetic brick “ OR “genetic circuit or genetic circuits “ OR
“genome-driven cell engineering” OR “genome engineering” OR “genome scale DNA
synthesis “ OR “homologous recombination” OR “industrial microorganism” OR
“integrated gene circuits” OR “I-isoleucine” OR “metabolic channelling” OR metabolic
engineering” OR “metabolic network” OR “microbial cell factory or factories” OR “minimal
genome” OR “model synthetic” OR “Molecular biotechnology “ OR “multi enzyme one pot”
OR “pathway engineering” OR “plasmid backbone” OR “platform to run” OR “probiotic”
OR “program verification” OR “protocell” OR “protocell creation” OR “recombinant gene
cluster” OR “Recombineering” OR “regulatory network” OR “stORardised genetic modular
part” OR “synthetic biology” OR “synthetic gene circuits” OR “stORardisation” OR
“synthetic DNA” OR “synthetic gene transcription” OR “synthetic genome” OR “synthetic
enzyme” OR “synthetic genomics” OR “synthetic metabolic pathway” OR “substrate
channelling” OR “top-down” OR “whole genome of a eukaryotic organism”
STEP 2: “Artificial nose” OR “Antioxidant” OR “Biosensor” OR “Emulsifier” OR “Enzyme”
OR “Feed” OR “Feedstock” OR “Fermentation product” OR “Flavour” OR “Flavour
enhancer” OR “Fragrance” OR “Food additive” OR “Food colour” OR “Food crop” OR
“food crop derived product” OR “Food ingredient” OR “Food preservative” OR “Gelling
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agent” OR “Health product” OR “Metabolite” OR “Nutraceutical” OR “Preservative” OR
“Probiotic” OR “Processing aid” OR “Stabiliser” OR “Sweetener” OR “Thickener”.
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Annex I-B: Deliverable 2 - Representative case studies of potential Synthetic Biology
food/feed applications and factors influencing their development and
commercialisation.
Representative case studies of potential Synthetic Biology food/feed
applications and factors influencing their development and
commercialisation
106
Contents
Item
1. Context: synthetic biology - what are the limits?
2. Updated inventory of synthetic biology food/feed products and
applications identified through literature search
3. Representative case studies
4.
5.
6.
7.
8.
106
Proposed classification of synthetic biology food/feed products and
applications
Potential societal responses of synthetic biology food/feed products and
applications
Conclusions
Next steps – planning Deliverable 3
Annexes
Page
2
6
17
23
25
30
31
38
Image from: http://undsci.berkeley.edu/article/0_0_0/howscienceworks_18
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1. Context: Synthetic biology - which are the limits?
Understandably our society is always interested in creating the means to identify and
implement quicker, more efficient, and less costly solutions to short- and long-term
challenges that we face. Synthetic biology is a field that offers futuristic products and
applications to address some of the crucial issues in relation to health and the environment
(e.g. green energy, ensuring food security for all, climate change, environmental pollution,
targeted health intervention) and quicker scientific progress in a number of fields107 (Khalil
& Collins, 2010; Schmidt, 2010; TNS-BMRB, 2010 - WP2; Bailey et al., 2012; Flari &
Chaudhry, 2012; Hoskisson, 2012; STOA workshop report, 2012). Synthetic biology seeks to
engineer new, or re-design exiting functions, in organisms, aiming to produce desired
products (e.g. drugs, food additives, biofuel, etc.), or a desired service (e.g. remediation)
(Heinmen & Panke, 2006; Alterovitz et al., 2009). Potential prospects for synthetic biology
appear to be vast, restrained perhaps by two major encompassing dimensions: the speed of
development of technological advances, and the degree of social acceptance of synthetic
biology products and applications. The latter is crucial as the term “synthetic biology”
implies non-natural processes in living organisms that are able to replicate and reproduce,
and hence an obvious technological association with organisms and products developed
through Genetic Modification (GM) and by synthetic biology. It would be sensible for one to
think that these two influencing dimensions could be positively correlated; i.e. the more a
technology and/or product/application are accepted by the public, the more possible that
resources (i.e. funds and time) would be allocated for its development and vice versa. The
moratorium on GM research in Europe could be a strong example of such correlation
(Amman, 2010; European Commission 2010).
Currently, certain technological areas or sub-fields of synthetic biology have been identified
(a review is shown in COGEM, 2013):

Synthetic genome (total or substantial parts of genome assembled through synthetic
DNA/RNA)
Factory of life: Synthetic Biologists reinvent nature with parts, circuits. Science news, January 12 2013. [Online]
Available at: https://www.sciencenews.org/article/factory-life [Last accessed 16 12/2013]
107
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
Metabolic pathway engineering

Regulatory circuits

Minimal genome organism

Orthogonal life

Protocell

Xenobiology
An outline of the above major technological areas or sub-fields of synthetic biology is shown
in Annex I. The analysis included in Annex I is based mainly on information published in
COGEM report 2013108.
Computational Biology (or Bioinformatics) contributes significantly to the technological
advances of synthetic biology as it allows researchers to model their ‘genetic circuits’ before
trying them out in live biological systems in order to choose the “fit for purpose” synthetic
genetic parts (Kuwahara et al., 2013). Additionally, recently developed modelling
techniques accommodate for the inherent variability in the biological systems (Tan and Liao,
2012; Toni and Tidor, 2013).
How close, however, are synthetic biology based organisms or products to reality? How
probable is it to design and create a synthetic, non-natural, organism that would be
engineered to deliver a desired product and/or a service and would be viable, and able to
replicate and reproduce? The outcome of a 15-year, highly costly but successful, experiment
run at the J. Craig Venter Institute in USA indicates that promises of Synthetic Biologists
may not be a fiction. Synthia, the first “synthetic organism” was announced in Science in
2010109. COGEM report in 2013110 reports that it took a team of 25 researchers 15 years to
complete the project, which cost more than 40 million US dollars. The future intention of the
team is to get on a further technological step and transplant the minimal genome they
108
COGEM (2013) Synthetic Biology – Update 2013. Anticipating developments in synthetic biology. [Online]
Available at: http://www.cogem.net/index.cfm/en/publications/publicatie/synthetic-biology-update-2013 [Last
accessed 16 12 2013]
109 Gibson DG, Glass JI, Lartigue C, Noskov VN, Chuang RY, Algire MA, Benders GA, Montague MG, Ma L,
Moodie MM, Merryman C, Vashee S, Krishnakumar R, Assad-Garcia N, Andrews-Pfannkoch C, Denisova EA,
Young L, Qi ZQ, Segall-Shapiro TH, Calvey CH, Parmar PP, Hutchison CA 3rd, Smith HO, Venter JC (2010 Jul 2;
Epub 2010 May 20). "Creation of a bacterial cell controlled by a chemically synthesized genome". Science 329
(5987): 52–6.
110 COGEM (2013) Synthetic Biology – Update 2013. Anticipating developments in Synthetic Biology. [Online]
Available at: http://www.cogem.net/index.cfm/en/publications/publicatie/synthetic-biology-update-2013 [Last
accessed 16 12 2013]
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identified for Mycoplasma genitalium into a cell to create Mycoplasma laboratorium, a synthetic
mycoplasma with the minimum genome needed to allow it to be viable and able to
replicate111.
The key question for this project is how probable it is that a synthetic biology food/feed
product will be on the market in the coming years? Particularly prospects of synthetic
biology for food and feed have been truly ambitious (Dos Santos, 2010; Ducat et al., 2012;
optimised crop production112), although most prominent research is focused on agricultural
applications. In the UK the vision of employing synthetic biology in optimised crop
production has been translated into a range of highly funded projects, e.g. projects that aim
to make non-leguminous crop plants able to fix nitrogen113. An alternative synthetic biology
approach using cyanobacteria within photosynthetic cells is being pursued by scientists at
Washington University in USA114.
A snapshot of broad opportunities of synthetic biology for food/feed products and
applications was given by Vitor Martins dos Santos in a meeting organised by the Health
and Consumers DG of the European Commission115. More recently, an inventory of existing
and possible products based on synthetic biology has been developed (and regularly
updated) by the synthetic biology project116, an initiative stemmed by the Woodrow Wilson
International Center for Scholars. The inventory (most recently updated on 27th of July 2012)
refers to certain groups of synthetic biology applications, i.e. biofuels, chemicals, food,
materials, medicines, other; the food sector is the least populated group this far, compared
with the other sectors (the listed applications are shown in Figure 1 below).
Information retrieved from: http://en.wikipedia.org/wiki/Mycoplasma_laboratorium
http://www.bbsrc.ac.uk/news/research-technologies/2012/121109-pr-investment-uk-synthetic-biologyresearch.aspx
113 For example Professor Giles Oldroyd at John Innes has received £2.5 million funding from BBSRC for research
in this area, (http://www.bbsrc.ac.uk/news/research-technologies/2012/121109-pr-investment-uk-syntheticbiology-research.aspx)
114 http://news.wustl.edu/news/Pages/25585.aspx
115 Synthetic biology in Food and Health. [Online] Available at:
http://ec.europa.eu/health/dialogue_collaboration/docs/ev_20100318_co08.pdf [Last accessed 16 12 2013]
116 http://www.synbioproject.org/
111
112
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Figure 1: Snapshot of the inventory of synthetic biology products – existing and possible (Draft – July
2013)117.
Our search for synthetic biology food/feed products and applications (existing or
forthcoming) investigated peer reviewed relevant publications, patents, grey literature,
reports from highly regarded organisations (e.g. European Commission, WHO, etc.) as well
as discussions within the project team. As described previously (in deliverable 1) the
identification of “pure” synthetic biology food/feed products applications was highly
complicated because of the large overlap with “older, more traditional” biotechnology
fields, most prominently GM. The latter was even more evident when searching patents
where it seems even more difficult to distinguish synthetic biology from long established
biotechnology fields, such as metabolic engineering, recombinant protein expression, etc.
(Rutz, 2009). The outcomes of this work therefore need to be interpreted with caution, and
as a “proof of concept” exploratory process.
117
Source: synthetic biology project:
http://www.synbioproject.org/process/assets/files/6631/_draft/synbio_applications_wwics.pdf
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2. Updated inventory of synthetic biology food/feed products and
applications identified through literature search
The final search strategy we developed in deliverable 1 (as shown in Table 3 in D1) was
employed to search available information sources (indicated in Table 2 in D1). This
resulted in identification of 713 articles and 108 patents as potentially relevant to the
objectives of the project. These were critically screened by the project team members to
identify the most relevant ones. The team applied the working understanding for
synthetic biology (as defined in page 6 in D1) in order to screen out products and
applications that were thought of as typically GM118.
Initial results119 from this exercise were enriched further (the protocol followed illustrated
in Figure 2 below), and an updated inventory of identified potential synthetic biology
food/feed products and applications is shown in Table 1.
Figure 2: The flow diagram illustrates the different literature search and screening phases followed
for the identification of the representative case studies for the project. Phase 1 was described in
detail in Deliverable 1 (D1 - pages 10 to 33). Human intelligence steps refer to the evaluations,
assessments, and elicitation of views from the project team members.
Some examples of such “rejected” products and applications are shown below in Table 4 in D1.
These included items that were considered as synthetic biology products and applications, items that refer
to products and applications that are based on methods which are rather closer to GM than synthetic biology,
albeit not strictly GM, and finally items that were considered as valuable because of the methodologies
described that would perhaps lead to the development of food / feed related synthetic biology products and
applications in the future (items are shown in detail in Table 5 in D1).
118
119
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1120
213
3
4
5
6
7
8
9
10
11
12121
13
14122
15
Aharoni, A. and E. Lewinsohn (2010). Genetic engineering of fruit flavors. Handbook of fruit and
vegetable flavors: 101-114. ISBN 978-0-470-22721-3
Akhila, A. (2007). Metabolic engineering of biosynthetic pathways leading to isoprenoids: Mono- and
sesquiterpenes in plastids and cytosol. Journal of Plant Interactions 2(4): 195-204.
Alchihab, M., J. Destain, et al. (2010). Production of aroma lactones by yeasts. Biotechnologie Agronomie
Societe Et Environnement 14(4): 681-691.
Azadi, P., N. V. Otang, et al. (2010). Metabolic engineering of Lilium x formolongi using multiple genes of
the carotenoid biosynthesis pathway. Plant Biotechnology Reports 4(4): 269-280.
Baeumchen, C., A. H. F. J. Roth, et al. (2007). D-mannitol production by resting state whole cell
biotransformation of D-fructose by heterologous mannitol and formate dehydrogenase gene expression
in Bacillus megaterium. Biotechnology Journal 2(11): 1408-1416.
Barghini, P., D. Di Gioia, et al. (2007). Vanillin production using metabolically engineered Escherichia coli
under non-growing conditions. Microbial Cell Factories 6. DOI 13 10.1186/1475-2859-6-13
Baumgartner, F., L. Seitz, et al. (2013). Construction of Escherichia coli strains with chromosomally
integrated expression cassettes for the synthesis of 2 '-fucosyllactose. Microbial Cell Factories 12. DOI 40
10.1186/1475-2859-12-40
Blankschien, M. D., J. M. Clomburg, et al. (2010). Metabolic engineering of Escherichia coli for the
production of succinate from glycerol. Metabolic Engineering 12(5): 409-419.
Bovy, A., E. Schijlen, et al. (2007). Metabolic engineering of flavonoids in tomato (Solanum lycopersicum):
the potential for metabolomics. Metabolomics 3(3): 399-412.
Brochado, A. R., C. Matos, et al. (2010). Improved vanillin production in baker's yeast through in silico
design. Microbial Cell Factories 9. 84
DOI 10.1186/1475-2859-9-84
Burgess, C., W. Sybesma, et al. (2003). Characterisation and improvement of riboflavin production in
Lactococcus lactis. Abstracts of the General Meeting of the American Society for Microbiology 103: O-088.
Burja, A. M. and H. Radianingtyas (2005). Marine microbial-derived nutraceuticals biotechnology: an
update. Food Science & Technology 19(1): 14-16.
Cimini, D., K. R. Patil, et al. (2005). Metabolic engineering of succinic acid production in Saccharomyces
cerevisiae. Journal of Biotechnology 118: S118-S118.
Cunningham, F. (2003). Metabolic engineering of pathways leading to fat-soluble vitamins and
antioxidants in plants: What have we learned and where are we going? Hortscience 38(5): 715-715.
Damude, H. G. and A. J. Kinney (2007). Metabolic engineering of seed oil biosynthetic pathways for
human health. Verpoorte, R., Alfermann, A. W., Johnson, T. S. Applications of Plant Metabolic
Engineering. 978-1-4020-6030-4(H). DOI 10.1007/978-1-4020-6031-1_10
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl.
fermentation product)
Health product
Gelling agent
Fragrance (relevant to
food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived
product
Colour agent
Additive
Article or patent
Antioxidant
Table 1: Updated and enriched inventory of items identified as relevant to the aims and objectives of the study. We were not able to source the articles highlighted in yellow.
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
120
Review
Concept paper
122
Abstract
121
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16
17
1813
19
2013
2113
22
23
24
25
26
27
28
29
30
3113
Dyer, J. M., D. C. Chapital, et al. (2003). Metabolic engineering of bakers' yeast for production of valueadded lipids. Abstracts of Papers American Chemical Society 225(1-2): 115-AGFD 115.
Field, D., M. Begley, et al. (2012). Bioengineered Nisin A Derivatives with Enhanced Activity against
Both Gram Positive and Gram Negative Pathogens. PLoS ONE 7(10).
Gaspar, P., A. R. Neves, et al. (2008). Engineering and re-routing of the metabolism of lactic acid bacteria.
Mayo, B., Lopez, P., Perez-Martinez, G. Molecular aspects of lactic acid bacteria for traditional and new
applications. ISBN 265-289. 978-81-308-0250-3
Han, X.-y., D.-q. Sun, et al. (2007). Gene regulation to lactic acid bacteria for increasing production of
flavor metabolite. Weishengwu Xuebao 47(6): 1105-1109.
Hendrawati, O., H. J. Woerdenbag, et al. (2010). Metabolic engineering strategies for the optimization of
medicinal and aromatic plants: realities and expectations. Zeitschrift Fur Arznei- & Gewurzpflanzen
15(3): 111-126.
Iijima, Y. (2010). Metabolic factory for flavors in fruits and vegetables. Handbook of fruit and vegetable
flavors: 705-727. ISBN 978-0-470-22721-3
Jin-Ho, C., R. Yeon-Woo, et al. (2005). Biotechnological production and applications of coenzyme Q10.
Applied Microbiology and Biotechnology 68(1): 9-15.
Jung, W. s., I.-M. Chung, et al. (2003). Manipulating isoflavone levels in plants. Journal of Plant
Biotechnology 5(3): 149-155.
Kim, S.-H., S.-Y. Kim, et al. (2009). Biosynthesis of Polyunsaturated Fatty Acids: Metabolic Engineering
in Plants. Journal of Applied Biological Chemistry 52(3): 93-102.
Kong M.K., Lee P.C. (2011). Metabolic engineering of menaquinone-8 pathway of Escherichia coli as a
microbial platform for vitamin K production.
Biotechnology and Bioengineering 108(8): 1997–2002
Koopman, F., J. Beekwilder, et al. (2012). De novo production of the flavonoid naringenin in engineered
Saccharomyces cerevisiae. Microbial Cell Factories 11. DOI 155 10.1186/1475-2859-11-155
Lee, J. H. and D. J. O'Sullivan (2008). Metabolic Engineering of Lactococcus lactis for the Development of a
One-Step Bioconversion of Lactose into Tagatose. Abstracts of the General Meeting of the American
Society for Microbiology 108: 473-473.
Lei, Z., W. Zinan, et al. (2007). Metabolic engineering of plant L-ascorbic acid biosynthesis: recent trends
and applications. Critical Reviews in Biotechnology 27(3): 173-182.
Lemuth, K., K. Steuer, et al. (2011). Engineering of a plasmid-free Escherichia coli strain for improved in
vivo biosynthesis of astaxanthin. Microbial Cell Factories 10. DOI 29 10.1186/1475-2859-10-29
Lewinsohn, E., R. Davidovich-Rikanati, et al. (2010). Functional genomics for the discovery of genes
affecting lemon basil aroma and their use in flavor engineering of tomato. ISHS Acta Horticulturae 860:
IV International Symposium on Breeding Research on Medicinal and Aromatic Plants - ISBMAP2009
Liu, N. and S. Zheng (2010). Advances in biological functions and biosynthesis regulation of tocotrienols.
Guangxi Zhiwu / Guihaia 30(1): 122-159.
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl.
fermentation product)
Health product
Gelling agent
Fragrance (relevant to
food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived
product
Colour agent
Antioxidant
Additive
Article or patent
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
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3213
33
34
3513
3613
37124
3813
39
40
41125
42
4313
44126
Lorence, A. and C. L. Nessler (2007). Pathway engineering of the plant vitamin C metabolic network.
Verpoorte, R., Alfermann, A. W., Johnson, T. S. Applications of Plant Metabolic Engineering: 1970217.
ISBN 978-1-4020-6030-4(H)
Lu, C.-H., J.-H. Choi, et al. (2011). Laboratory-Scale Production of Tomato Carotenoids Using
Bioengineered Escherichia coli. FASEB Journal 25.
Luo, H., J. Song, et al. (2013). Cloning and expression analysis of a key device of HMGR gene involved in
ginsenoside biosynthesis of Panax ginseng via synthetic biology approach. Acta Pharmaceutica Sinica
48(2): 219-227.
Mattoo, A. K., V. Shukla, et al. (2010). Genetic Engineering to Enhance Crop-Based Phytonutrients
(Nutraceuticals) to Alleviate Diet-Related Diseases. Bio-Farms for Nutraceuticals: Functional Food and
Safety Control by Biosensors. M. T. Giardi, G. Rea and B. Berra. 698: 122-143.
Mehrotra, S., L. U. Rahman, et al. (2010). An extensive case study of hairy-root cultures for enhanced
secondary-metabolite production through metabolic-pathway engineering. Biotechnology and Applied
Biochemistry 56: 161-172.
Melzer, G., M. E. Esfandabadi, et al. (2009). "Flux Design: In silico design of cell factories based on
correlation of pathway fluxes to desired properties. Bmc Systems Biology 3: 120
Misawa, N. (2009). Pathway engineering of plants toward astaxanthin production. Plant Biotechnology
26(1): 93-99.
Moldrup, M. E., +F. Geu-Flores, et al. (2011). Modulation of sulfur metabolism enables efficient
glucosinolate engineering. BMC Biotechnology 11(12): (31 January 2011)-(2031 January 2011).
Park, J. H. and S. Y. Lee (2010). Metabolic pathways and fermentative production of L-aspartate family
amino acids. Biotechnology Journal 5(6): 560-577.
Peiru, S., E. Rodriguez, et al. (2008). Metabolically engineered Escherichia coli for efficient production of
glycosylated natural products. Microbial Biotechnology 1(6): 476-486.
Reuben, S., L. J. Cseke, et al. (2006). Molecular biology of plant natural products. Cseke, L. J., Kirakosyan,
A., Kaufman, P. B., Warber, S. L., Duke, J. A., Brielmann, H. L. Natural products from plants: 165-202.
ISBN 0-8493-2976-0\978-0-8493-2976-0
Sato, F. and K. Matsui (2012). Engineering the biosynthesis of low molecular weight metabolites for
quality traits (essential nutrients, health-promoting phytochemicals, volatiles, and aroma compounds).
Altman, A, Hasegawa, P. M., Plant Biotechnology and Agriculture: Prospects for the 21st Century, 443461. 978-0-12-381467-8.
So-Yeon, S., H. Nam Soo, et al. (2011). Production of resveratrol from p-coumaric acid in recombinant
Saccharomyces cerevisiae expressing 4-coumarate:coenzyme A ligase and stilbene synthase genes.
Enzyme and Microbial Technology 48(1): 48-53.
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl.
fermentation product)
Health product
Gelling agent
Fragrance (relevant to
food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived
product
Colour agent
Antioxidant
Additive
Article or patent
x
x
123
x
x
x
x
x
x
x
x
x
x
x
x
x
In the sense that vitamin C that is enhanced in a crop is a crucial supplement for human diet.
Method paper; description of process / strategy to optimize strain and bioprocess.
125 Polyketides are pharmacologically important. The paper is a method paper for demonstrating a technique for efficient production of glycosylated natural products. The evaluation of this paper concentrates on how the methods described could be used in order
to produce food/feed SynBio products/applications.
126 Production of resveratrol, a polyphenolic compound.
123
124
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45127
46
47
48
48b
49128
50
51
52
53129
54
5513
56
5713
58
Sun-Young, Kang, C. Oksik, et al. (2012). Artificial biosynthesis of phenylpropanoic acids in a tyrosine
overproducing Escherichia coli strain. Microbial Cell Factories 11(Dec.): 153-153.
Thanasomboon, R., D. Waraho, et al. (2012). Construction of synthetic Escherichia coli producing slinalool. Proceedings of the 3rd International Conference on Computational Systems-Biology and
Bioinformatics. J. H. Chan, A. Meechai and C. K. Kwoh. 11: 88-95.
Van der Straeten et al. (2010) Patent Application Publication US 2010/0183750 A1: Fortification of plants
with folates by metabolic engineering.
Wang, Y., H. Chen, et al. (2010). Metabolic engineering of resveratrol and other longevity boosting
compounds. Biofactors 36(5): 394-400.
Jeandet et al., 2013: Metabolic Engineering of Yeast and Plants for the Production of the Biologically
Active Hydroxystilbene, Resveratrol. J Biomed Biotechnol. 2012; 2012: 579089. Published online 2012
May 13. doi: 10.1155/2012/579089
Watts, K. T., P. C. Lee, et al. (2006). Biosynthesis of plant-specific stilbene polyketides in metabolically
engineered Escherichia coli. BMC Biotechnology 6(22): (21 March 2006)-(2021 March 2006).
Won-Heong, Lee., P. Panchalee, et al. (2012). Whole cell biosynthesis of a functional oligosaccharide, 2′ fucosyllactose, using engineered Escherichia coli. Microbial Cell Factories 11(April): 48-48.
Yu, O. (2009). Genetic regulation and metabolic engineering of isoflavone biosynthesis. Abstracts of
Papers of the American Chemical Society 238: 294-294.
Yue, A.-Q. and R.-Z. Li (2009). Metabolic Engineering to Produce Novel Renewable Industrial Fatty
Acids in Oilseeds. Zhongguo Shengwu Huaxue yu Fenzi Shengwu Xuebao 25(6): 501-509.
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Hugenholz et al. (2002). Metabolic engineering of lactic acid bacteria for the production of
Nutraceuticals. Antonie van Leeuwenhoek 82: 217–235, 2002.
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strains and foodborne pathogens and potential implications of engineered probiotic intervention in
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10.1021/jf4007104
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Patent 0080233622
x
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Nutraceutical
Metabolite
Ingredient (incl.
fermentation product)
Health product
Gelling agent
Fragrance (relevant to
food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived
product
Colour agent
Antioxidant
Additive
Article or patent
x
x
x
x
x
x
x
x
x
x
x
Production of phenylpropanoic acids that draws increasing attention due to pharmaceutical properties. Method paper that could be used as an example for production of SynBio food/feed products/applications. Use of
phenylpropanoic acid in food industry is reported: http://en.wikipedia.org/wiki/Phenylpropanoic_acid
127
128
129
Synthesis of plant-specific stilbene polyketides with which numerous health benefits are associated.
Background method paper that could facilitate metabolic engineering of fragrances in the future.
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59
60
61130
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Kayser, O. (2010). Metabolic engineering strategies for the optimization of medicinal and aromatic
plants: expectations and realities. Acta Horticulturae. D. Baricevic, J. Novak and F. Pank: 199-204.
Labuda, I. (2011). Biotechnology of vanillin: vanillin from microbial sources. Havkin-Frenkel, D.,
Belanger, F. C., Handbook of vanilla science and technology: 301-331. ISBN 978-1-4051-9325-2
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Oke, M. and G. Paliyath (2010). Flavor from transgenic vegetables. Handbook of fruit and vegetable
flavors: 681-692. ISBN 978-0-470-22721-3
Rito-Palomares, M. (2010). Bioengineering strategies for the purification of biological products. Journal
of Biotechnology 150: S357-S357.
Schwab, W., T. Hoffmann, et al. (2010). Metabolic Engineering in Fragaria x ananassa for the Production
of Epiafzelechin and Phenylpropenoids. Flavor and Health Benefits of Small Fruits. M. C. Qian and A.
M. Rimando. 1035: 293-300.
69
Sun-Hwa, Ha., L. Ying Shi, et al. (2010). Application of two bicistronic systems involving 2A and IRES
sequences to the biosynthesis of carotenoids in rice endosperm. Plant Biotechnology Journal 8(8): 928938.
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7(3): 343-353.
Wakasa, K. and A. Ishihara (2009). Metabolic engineering of the tryptophan and phenylalanine
biosynthetic pathways in rice. Plant Biotechnology 26(5): 523-533.
71
130
131
Supplement
Sweetener
Stabiliser
Processing aid
Probiotic
Preservative
Metabolite
Ingredient (incl.
fermentation product)
Health product
Gelling agent
Fragrance (relevant to
food flavouring)
Flavour enhancer
Flavouring agent
Feed - Feedstock
Enzyme
Emulsifier
Crop or Crop derived
product
Nutraceutical
x
Farasat, I., J. Collens, et al. (2011). Efficient optimization of synthetic metabolic pathways with the RBS
Library Calculator. Abstracts of Papers American Chemical Society 241: 161-BIOT.
6413
6714
x
Diretto, G., N. Schauer, et al. (2010). Systems biology characterization of engineered tomato fruits with
improved carotenoid content provides novel insights on the interplay between pigment biosynthesis and
post-harvest characteristics. Journal of Biotechnology 150: S548-S549.
Itkin, M. and A. Aharoni (2009). Bioengineering. Osbourn, A. E., Lanzotti, V. Plant-Derived Natural
Products: Synthesis, Function, and Application: 4350473. ISBN 978-0-387-85497-7
6613
Colour agent
Abbas, C. A. and A. A. Sibirny (2011). Genetic control of biosynthesis and transport of riboflavin and
flavin nucleotides and construction of robust biotechnological producers. Microbiology and Molecular
Biology Reviews 75(2): 321-360.
Alper, H. S. and C. Wittmann (2013). Editorial: How multiplexed tools and approaches speed up the
progress of metabolic engineering. Biotechnology Journal 8(5): 506-507.
6313
65
Antioxidant
Additive
Article or patent
x
x
x
x
x
x
Method paper on computational biology.
Method paper for modelling parameters for designing strains of Escherichia coli.
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The products/ applications included in the updated inventory were evaluated in
regard to the main technological areas concerned. No food/feed product or
application that was based on synthetic biology in terms of either regulatory circuits,
minimal genome organisms, orthogonal life, protocells, or xenobiology was
identified. Therefore majority of the products/applications included in Table 1 is
concerned with products or future applications that appeared to be based on either
DNA synthesis/ synthetic genome, and/or metabolic engineering.
The project team members evaluated further each of the identified items in regard to
the following dimensions:

Technological area or sub-field of synthetic biology, including the number of
genes involved.

Modified organism/s concerned, i.e. transgenic plant; bacteria; yeast.

Products and applications concerned; if not current, how far from being
considered at the market?

Risks: health, environment, socioeconomic, including uncertainties – what are
we most uncertain about?

Benefits: health, environment, socioeconomic.

Costs – anticipated total cost and stakeholders sharing the cost, e.g. academia,
industry, government.
As stated in Deliverable 1, this far, synthetic biology is not defined by one universal
definition across the world (see Table 1 in D1). What is more, the existence of
synthetic biology as a distinct area of research and/or technology is being disputed.
To our knowledge, the term Synthetic biology is being used to cover a number of
fundamentally different approaches, in terms of methods and materials used,
epistemic approach and key actors involved (O’Malley et al., 2007). The definition
given on page 2 of D1 is one that incorporates the concepts of engineering as its
“raison d'etre”, or otherwise, the purpose and usefulness of the synthetic organism.
Researchers who follow this engineering approach stress that the key to synthetic
biology is the application of engineering principles (such as modularisation,
standardisation, characterisation), and the implementation of systematic design
(Endy, 2005; Kitney and Freemont, 2012). As such, they are adamant that synthetic
biology “is not a direct extension of genetic engineering” (Kitney and Freemont, 2012,
p.2035).
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In the present work we tried to follow a pragmatic approach to identify Synthetic
Biology food/feed products and applications. Although such pragmatic approach is
crucial in order to perform this kind of study, it is also important to acknowledge that
debates about “what is synthetic biology exactly?” and even more often about “whether
synthetic biology differs from GM, and if so how?” do exist and are likely to persist for the
foreseeable future. One thing that perhaps was not defined very precisely in our
pragmatic definition was the qualitative, engineering aspect of synthetic biology
outputs. Our working definition (Table 1 in D1) appeared focused on a quantitative,
rather than on a qualitative distinction; nevertheless, we need to emphasize that the
underlying rationale of the project team members was equally focused on the
engineering aspect of novel, synthetic (i.e. artificial) organisms, and in particular on
“how much of the produced organism was designed as new”? When thinking about
products and applications where existing (e.g. whole metabolic pathway of a plant)
or artificial (partial of full) genome (e.g. xenobiology) is engineered into an existing
cell (e.g. recombinant E.coli) one needs to consider that the end product would carry a
combined genome from two different sources, e.g. then the results is a bacterial strain
which in Xth percent characteristic is also a plant. This will inevitably increase
complexity of regulation of future permutations between disparate species and
totally synthetic pathways and whole organisms.
Given the above, the project team members agreed upon criteria to employ in order
to evaluate the identified items in the updated inventory and assess how close each
one is to the concept of synthetic biology (shown in Box 1 below).
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Box 1: Criteria employed by the project team members in order to screen the items that were assessed as
being most closely to the concept of synthetic biology.
Close to the concept of
Synthetic Biology and will
be assessed further
Whole biosynthetic pathway is
concerned and there is production of
small molecules.

Outcome is of artificial concept, i.e. it
does not exist in nature.

Far from the concept of
synthetic biology and will not
be assessed further
Just mutation

Overexpression

Gene silencing

Gene product is a protein or a
peptide

One or few genes in a metabolic
engineering pathway are concerned

This evaluation phase allowed project team to map the identified items (n=71) against
two subjective scales: (a) a scale to describe how close each item is to any of the
concepts of synthetic biology (results shown in Figure 3 below), and (b) a subjective
timeline of product development that was designed to describe how close each item
is thought to be to being on the shelf or in commercial production (results shown in
Figure 4 below).
During this evaluation process, the project team members excluded three papers
from any further assessment (i.e. items 28, 41 and 46 were considered out of scope).
Additionally, project team members considered a subset of review articles for further
evaluation during the next phase of the project132.
132
These review papers are: 1, 2, 18, 20, 21, 31, 32, 35, 36, 38, 42, 43, 55, 57, 63, 64, 70
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Figure 3: The identified items were evaluated by the project team members in terms of how close each
of these are to the concept of synthetic biology. Project team members took into account a number of
criteria, including the number of genes involved, whether an engineering concept was evident, as
well as the criteria listed above in Box 1. The initial (and furthest from the concept of synthetic
biology) point was set as a product in which one or very few functional units were modified, e.g.
transgenic herbicide resistant oilseed rape. The point closest to the concept of synthetic biology was
set as a new, totally artificial probiotic organism that would be able to survive and reproduce and
would be designed and engineered to produce an entirely synthetic probiotic.
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Figure 4: Project team members evaluated the identified items against a subjective timeline of product
development. Four stages were introduced in the scale: (a) conceptual, (b) a phase where parts of the
product have been produced in the lab, (c) the product has been produced in the lab, and (d) product
in on shelf or has been produced in the field.
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3. Representative case studies
Following the evaluation of identified items in terms of how close each item is to (a)
the concept of synthetic biology (results shown in Fig. 3 above), and (b) being
marketed (results shown in Fig. 4 above), the project team decided to assess in more
detail the items that were mapped closer to the concept of synthetic biology, i.e.
items mapped beyond point “5” in the subjective scale drawn by the project team
members (Fig. 3 above). Thus, the following products and applications were
considered in more detail:

Synthesis of vanillin in engineered E. coli (No 6133 in Table 1) or yeast (S.
cerevisiae; No 10 in Table 1).

Synthesis of coenzyme Q10 (No 22 in Table 1).

Production of vitamin K (No 25 in Table 1).

Synthesis of metabolites via the phenylpropanoid and isoprenoid pathways
(several end products could be concerned; No 45 in Table 1).

Engineering of probiotic bacteria (i.e. Lactococcus lactis; No 55 in Table 1), again
with several possibilities.
A more detailed evaluation of these selected products and applications is shown in
Table 2 below. Project team members are further developing each of these selected
products and applications as detailed case studies to employ in assessment of
whether the existing regulatory schemes are “fit for purpose” in the next phase of
the project (see below section 7).
In addition to the above selected products and applications the team decided to
include (a) one more application in the representative case studies, based on existing
This is a product that was considered quite close to marketing; http://www.evolva.com/products/vanilla ;
http://www.evolva.com/media/press-releases/2013/2/5/iff-and-evolva-enter-pre-production-phase-naturalvanillin-global-food
133
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research that seeks to make non-leguminous plants able to fix atmospheric
nitrogen134, and (b) possibly a synthetic yeast135.
134
135
http://www.genomeweb.com/us-uk-provide-12m-nitrogen-fixing-bacteria-and-crop-studies
http://syntheticyeast.org/
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Table 2: Evaluation of selected products and applications as case studies to assess the “fit for purpose” of existing regulatory schemes in the
next phase of the project.
136
No
Product / Application
Health risks
Environment risks
Socioeconomic risks
Benefits
6,10
Synthesis of vanillin in
engineered E. coli or yeast
(S. cerevisiae)
Possible, as whole metabolite profile
of yeast would be consumed. SynBio
yeast may also produce some
harmful substances as a by-product.
Unknown
Depending on whether
is labelled as
“natural”137.
Possible environmental
benefits if bacteria (in
case E. coli is concerned)
are used to use waste
materials as energy
source.
Plant products are not “alone” in
terms of giving natural flavour;
accompanied by a number of other
compounds.
Impacts (positive
and/or negative) on
local biodiversity in
areas where vanilla is
grown as a crop.
E.coli: none as the product will be
extracted and quality assessed for
purity.
economic, social and
cultural impacts on
producers of vanilla
from agricultural crops
Local impacts on
biodiversity in areas
where vanilla is gown as
a crop.
However, amount of
waste that could be
consumed would be
very small. It is also
unlikely that
manufacturers would
design a yeast that
grows on waste in this
case (for a high value
compound), as they
want to be able to
control the growth
conditions very
precisely.
If accepted as safe, a
yeast that also generates
‘natural’ vanilla flavour
in situ will be highly
136
137
Numbers as cited in Table 1 above.
www.foe.org/projects/food-and-technology/synthetic-biology/No-Synbio-Vanilla
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No
136
Product / Application
Health risks
Environment risks
Socioeconomic risks
Benefits
sought after.
22
Synthesis of coenzyme
Q10 (vehicle: E.coli)
None was identified during
evaluation as the product will be
extracted and quality assessed for
purity.
Unknown
Current production of
Q10 very expensive.
Already this is
consumed as a
“supplement”; possibly
would be more
accessible (as it would
be less expensive to
produce). Nevertheless,
it should be emphasized
that the product fits
with a particular
concept of health, based
on consuming
individual ‘beneficial’
molecules rather than
having a healthy mixed
diet and lifestyle, and
more broadly
acknowledging the
social dimensions of
health.
25
Production of vitamin K
(vehicle: E.coli)
None was identified during
evaluation as the product will be
extracted and quality assessed for
purity.
Unknown
Unknown
Current production via
fermentation; possibly
of high cost.
If the synthetic biology
product is grown on
waste then
environmental benefits
would be included.
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136
No
Product / Application
Health risks
Environment risks
Socioeconomic risks
Benefits
45
Synthesis of metabolites
via the phenylpropanoid
and isoprenoid
pathways(vehicle: E.coli)
Unknown
Unknown
Depends on how the
product would
“compete” with
conventional production
of the end products (e.g.
caffeine).
Sustainable production
of products of interest
could be a benefit.
Engineering of probiotic
bacteria (Lactococcus
lactii).
A “live” organism that would be
consumed. Could be included in a
number of end-products. If anything
is going wrong in the design and the
output has unknown effects (until
they get recorded and monitored –
this maybe particularly true when
chronic effects are concerned). This
possibility should increase
complexity of regulation.
Is this something that
people would buy “of
the shelf”?
Depends on the
particular probiotics
“produced”.
55
This could be considered
as an “umbrella”
platform being a
probiotic bacteria.
Unknown risks.
Unknown.
When we sell or eat
something that is alive
(e.g. probiotics) many
unknowns exist:

Where does it
survive?

For how long
does it
survive?
If the synthetic biology
product is grown on
waste then
environmental benefits
would be included (see
above). However, as
mentioned above highvalue/low-volume
products will ever be
produced using yeast
that grows on waste.
That scenario is more
likely for high
volume/low value
products such as
biofuels.
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No
136
Product / Application
Health risks
Complications may arise if point of
purchase and point of consumption
differ.
Environment risks

Socioeconomic risks
Benefits
Which are any
interactions
with the
environment?
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4. Proposed classification of synthetic biology food/feed products and
applications
The work conducted in the project so far also aimed to classify potential synthetic biology
food and feed products according to types of applications; whether this would be
appropriate for identifying gaps in the existing regulatory frameworks remains to be seen
during the next phase of the project. Inevitably however, societal responses will need to be
addressed in any classification scheme, and including regulatory frameworks. In particular
there are two dimensions that the project team believes would be crucial for discussions of
societal responses to such products:

Contained use and deliberate release138
In European and UK regulations, a distinction is made between applications involving
genetically modified micro-organisms that are intended for “contained use” in industrial
fermenters for the production of compounds such as additives, colourings, emulsifiers,
flavourings and those intended for “deliberate release into the environment”, such as plant
crops and micro-organisms used as pesticides, fertilisers or other kinds of in-field growth
promoters. Almost all of the applications listed in Table 1 would fall into the “contained use”
category, and this is expected to have important implications for regulation and societal
responses.

Classification by type of benefit
The second dimension is the types of benefit, which will be absolutely crucial in shaping
societal responses: who is supposed to benefit? In what way? What existing
product/technology would this replace? In what way/s will it be better that the existing
product/technology: would it be cheaper, faster, healthier, more environmentally
sustainable, fairer? Possibilities could include:
o Improving crop yield.
o Producing food (including supplements, nutraceuticals) more sustainably.
o Producing processed food/feed more cheaply or efficiently.
138
Marris and Jefferson (2013)
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o Increasing nutritional properties of food/feed; improving the flavour or
appearance of food.
o Improving nutritional properties of food to alleviate or prevent diet-related
diseases for wealthy consumers (mostly in the global North).
o Improving nutritional properties of food to alleviate or prevent diet-related
diseases for poor consumers (mostly in the global South).
o Producing food with pharmaceutical (or ‘nutraceutical’) properties to alleviate or
prevent non-diet related illnesses or improve health.
One could look into this aspect from the perspective of ‘enabling’ platforms; platforms that
can be used to develop many products (possibly affecting a number of sectors. To our
knowledge such platforms could refer to either (a) a particular pathway, or (b) a
microorganism (e.g. a probiotic organism) or yeast139.
An example for the former is the plant isoprenoid pathway; many agronomical and
biotechnological applications may be based on the plant isoprenoid metabolism (Vranova et
al., 2012), thus providing the possibility for a plethora of synthetic biology end products in a
number of sectors (e.g. biofuels, pharmaceuticals, etc.). The engineering of such broad
pathways would ensure speed of development of relevant end products. Nevertheless, they
may harbour monopolies that in time could hamper innovation and collaborations (EASAC,
2011). An example of the latter exist in one of our case studies, in particular the probiotic
bacteria (No 55 in Table 2 above).
139
http://syntheticyeast.org/
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5. Potential societal responses of synthetic biology food/feed products and
applications
There has been some research on public attitudes to synthetic biology already, using both
quantitative and qualitative research methods. The results are consistent with those from
more than three decades of social science research on public responses to technological risks,
and to emerging fields of science and technology, notably agricultural GMOs, and important
lessons can be learnt from that previous research.
Quantitative survey research provides relatively superficial data on public responses to
science and technology which needs to be interpreted with caution. Deeper and more
nuanced understandings of public responses are obtained from qualitative research, for
example using focus groups.
Social science research on public responses (especially from the field of “science and
technology studies”, or STS) reveals that members of the lay public (meaning citizens with
no direct involvement in the field in question), when they express themselves in focus
groups, do not express straightforward “for” or “against” views about any particular field of
science and technology. These focus group participants tend to be more nuanced and
ambivalent than is portrayed in “folk theories” (Rip, 2006) or “myths” (Marris, 2001) that
pervade the narratives of scientists and science policy actors. For synthetic biology, these
more nuanced positions are apparent from the workshops held with members of the public
for the BBSRC/EPSRC dialogue (Bhattachary et al., 2010) (see Annex 2), and in focus groups
conducted by Pauwels in the USA (Pauwels 2009, 2013).
Some key messages relevant to synthetic biology are:
Applications matter. There are certain key factors that need to be taken into account:

The intended purpose(s) of an application, including the extent to which claims
about benefits are credible.

The distribution of anticipated risks and benefits.

The extent to which those exposed to potential risks will be informed, and will have
a choice about their exposure.
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
Whether or not the intended use of the application will entail the deliberate release
into the environment of a live genetically engineered organism.
The often-heard notion that members of the public “accept” medical applications of GMOs
but “reject” food and agricultural GMOs, and that this is due to a preference for applications
that deliver direct benefits, is true to some extent, but is too simplistic. Differences in
responses to medical and food applications of GMOs are affected by a number of other
important factors including: whether the GMO will be released into the environment, how
risks and benefits are distributed, the length and regularity of exposure to the risks, how
information - and in particular uncertainties - is communicated and by whom, the extent to
which those exposed to risks have any choice about their exposure; the extent to which
unintended negative effects can be reversed if they arise; and judgments about differences in
the regulations in place in the two sectors (see further details below).
Applications that require the release of GM crops or bacteria in the environment will be
appraised very differently from those that rely on the contained use of bacterial or yeast cells
in industrial fermenters; and this applies equally to medical and agri/food applications.
Members of the public tend, unsurprisingly, to express more positive views towards
applications that are presented as achieving health, environmental or other societal benefits.
However, and crucially, members of the public will, given the chance, examine claims about benefits
and make judgements about their credibility that will affect their appraisal of specific applications.
Thus, claims about benefits will be examined for evidence that they will *actually* be delivered, in the
relevant local context(s). Claims that do not fit well with peoples’ experience of the behaviour
of institutions will, in particular, be treated with scepticism. Claims about benefits that are at
odds with what is actually ultimately delivered can generate further scepticism and fuel
public controversy (e.g. promises to feed the world’s poor compared to delivering herbicide
tolerant soya beans used to feed farm animals and in processed food consumed by wealthy
populations in the Global North). Such examination is more likely to occur (and/or be
evident) in focus groups and real life situations than in the context of a questionnaire survey
(The only way questionnaire respondents can express scepticism is to refuse to answer a
question, and indeed “don’t know”responses are particularly high in surveys of public
attitudes to science and technology).
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The importance of moving away from “synbiophobia-phobia”
Synthetic biology distinguishes itself from other fields by the attention it has paid to public
views at a stage when the field is still emerging and few, if any, products/applications have
been developed. Nanotechnology is probably the only other example where this occurred
(in the first decade of the 2000s). In both these cases, these initiatives are largely driven by
fears, among promoters of the field, of negative reactions from members of the public, with
the ‘GM debacle’ over GM crops and foods regularly cited as the example of a disastrous
situation that we need to prevent from re-occurring and must ‘learn lessons from’. With
respect to nanotechnology, Rip (2006) argued that the GM case contributed to a
“nanophobia-phobia - the phobia that there is a public phobia"; and reflecting on how
synthetic biology and nanotechnology are both defined as 'convergent' technologies,
Torgersen wryly commented: "”convergence” may have an additional meaning: European
technology developers seem to converge in their fear that the public might react negatively"
(Torgersen, 2009).
Discussions about synthetic biology are permeated with what one may call “synbiophobiaphobia”, as repeatedly demonstrated, for example, during panel discussions at the SB6.0
conference (the premier international conference on synthetic biology) in London in July
2013, and the way this was reported in the media (e.g. (Shukman 2013). With respect to
nanotechnology, Rip further argued that “the concern of nanoscientists and technologists
about public concerns (painted as phobia about nano) drives their views, rather than actual
data about public views” (Rip, 2006). It is crucial for synthetic biology not to make the same
mistake. In this section, we therefore review and critically analyse data about public views to
synthetic biology, and relevant other research on public views to other emerging
technologies, notably agricultural GMOs.
Factors that influence different public and societal responses to medical and agri/food
applications of biotechnology
Survey data that returns more positive attitudes to medical GMOs than to agri/food GMOs
is often interpreted as meaning that members of the public are “more accepting” of
applications where they can see direct benefits to themselves or those close to them. This is
110 | P a g e
often further qualified as an egotistical position, and an irrational one since people seem
willing to ignore potential risks in one area and not in another, even though “the
technology” involved (modification of DNA) is the same. Such interpretations are
challenged by qualitative data from focus groups, which reveals that members of the public
express positive views towards applications that have intended benefits for society in
general (e.g. for more sustainable agricultural practices), and/or for populations outside of
their immediate social circle, including populations in the global South. Additional factors
‘complexify’ these responses and help to explain the generally observed difference, in
questionnaire surveys, between “red” (medical) and “green” (agricultural) applications of
biotechnology, in particular:

Living GM organisms used for medical applications are generally “contained” rather
than “released”, and when medical applications involves the “release” of crops or
insects public views are less positive.

The benefits and risks of medical applications accrue to the same populations,
whereas the benefits and risks of food applications are unequally distributed:
benefits to seed companies, farmers, food manufacturers (sometimes also possibly to
the environment) and risks to the general population and to the environment.

Patients are given detailed, personalised information by people they trust (doctors)
about the risks and benefits entailed, and this includes candid information about
uncertainties, including “unknown unknowns”, i.e. drug information leaflets list
possible side effects and also ask patients to report any unanticipated unlisted effects
to their doctors.

Having received all this information, patients and their families can make their own
decisions about whether or not to use a proposed therapy.

Risk regulations for medicines and clinical procedures are more well-known and
trusted than those for environmental risks.

Corrective action can be taken: a patient can stop taking the medicine, medicines
(such as thalidomide) can be withdrawn from the market if problems become
apparent.
Moreover, medical applications do not receive blanket approval from focus group
participants. Research reveals that members of the public do express concerns also about
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medical applications, for example about whether innovation is overly driven by the interests
of big firms, and how scientific advances can affect our society in terms of increasing
inequalities between rich and poor, and encouraging trends towards greater ‘consumer
culture’ (e.g. see table on p.26 of UK Synbio Dialogue Report). And responses to medical
applications of synthetic biology that require the release of live organisms into the
environment (e.g. “sterile” mosquitoes) are not overwhelmingly positive (see Box 2 below),
and this is confirmed by on-going public controversies surrounding experimental releases of
these mosquitoes in Brazil, the Cayman Islands and Florida.
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6. Conclusions

Our literature search has not shown any pure synthetic biology food/feed products
applications.

Nevertheless, we have identified clear trends that indicate what types of synthetic
biology products and applications in the food/feed sectors could potentially be
developed in the future140.

In view of these results, we have identified a number of representative case studies to
illustrate hypothetical synthetic biology food/feed products to present in detail to
experts, risk assessors and regulators so that any potential inadequacies and gaps in
the regulatory schemes and frameworks can be proactively identified ahead of time.
For example:
o
Vanillin synthesised by synbio bacteria;
o
Yeast that can generate vanilla (and potentially other) flavours in food
products in situ;
o
Coenzyme Q10 synthesised by synbio microbes;
o
Vitamin K synthesised by synbio microbes;
o
Industrial chemicals synthesised via phenylpropanoid or isoprenoid
pathways by synbio microbes;
o
Enhanced levels, and potentially a multitude of supplements (different
vitamins, antioxidants, etc) by a single strain of synbio probiotic bacteria.
o
Non-leguminous crops able to fix atmospheric nitrogen141 - reducing the need
for synthetic fertilisers;
o
Synbio microbes to produce new industrial feedstock compounds142.
We believe this approach would be extremely valuable as it would allow regulators to
prepare in advance for a “fit for purpose” regulation of such products. This in turn is
anticipated to increase transparency and trust in regulatory processes, and also facilitate safe
development of the new technology in this important sector of economic growth.
140
Some more advanced developments may exist in the area of biosensors but the project team members
considered these as out of scope for this work.
141 http://www.genomeweb.com/us-uk-provide-12m-nitrogen-fixing-bacteria-and-crop-studies
142 http://syntheticyeast.org/
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7. Next steps – planning Deliverable 3
During the next phase of the project the work will focus on reviewing the regulatory
frameworks currently in place in terms of “fitness for purpose” to regulate synthetic biology
food/feed products/applications.
The project team members will prepare detailed case studies based on the selected products
and applications (see section 4 above), and in collaboration with the customer will identify
regulatory frameworks that could potentially be appropriate for regulating such products
and applications. For example, the novel food regulations, GM regulations (Directive
2001/18, regulation 1829/2003, GMO Deliberate Release Regulations 2002); international
guidance that would be beneficial to be taken into account when regulating such products,
(e.g. Guidelines for the Appropriate Risk Governance of Synthetic Biology (IRGC, 2010143),
WHO Scientific Working Group on Life Sciences Research and Global Health Security144,
etc.). The identified regulatory and/or guidance documents will be screened by project team
members in order to prepare the platforms for discussion with regulators during a
facilitated meeting/workshop.
Following the review of potential regulatory frameworks and/or guidance the project team
will organize and host a one day meeting in which a small number of experts and major
stakeholders will be invited to attend and contribute to the meeting. These may include FSA
regulators; Prof Rob Edwards from Fera/University of York; Academia (e.g. Imperial
College, UK); Dr Philippe Martin (European Commission - DG Sanco); a representative from
the UK Advisory Committee on Releases to the Environment (ACRE); Defra GM team;
Industry (e.g. Food and Drinks Federation), NGOs (e.g. Greenpeace International Science
Unit – Exeter University), etc. The full list of stakeholder participants will be discussed with
all project members and social science experts, and it will be agreed upon with FSA during
the 2nd project meeting which is scheduled to occur in project month 3.
International Risk Governance Council (2010) Guidelines for the Appropriate Risk Governance of synthetic
biology [Online] Available at: http://www.irgc.org/IMG/pdf/irgc_SB_final_07jan_web.pdf [Last accessed on
11/09/2012]
144 WHO Scientific Working Group on Life Sciences Research and Global Health Security [Online] Available at:
http://www.who.int/csr/resources/publications/deliberate/WHO_CDS_EPR_2007_4n.pdf [Last accessed on
11/09/2012]
143
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The aim of the meeting would be to present to the regulators and invited experts the
selected case studies with the view to (a) identifying the gaps in the existent regulatory
frameworks for regulating current and forthcoming synthetic biology food/feed
products/applications (outcome of objective 1), and (b) discussing possible approaches to
address the identified gaps, e.g. prioritizing research directions, possibilities for
international harmonization of regulatory frameworks, etc.
Following the completion of the facilitated meeting the project team will compile a draft
report to include:

A summary of outputs already included in Deliverables 1 and 2

The findings from the meeting, i.e. identified gaps in terms of “fitness of purpose” of
current regulatory frameworks to regulate such products

Recommendations for potential approaches to address the identified gaps

Recommendations for feasible and appropriate implementation mechanisms
The draft report will be finalized, presented and discussed in detail with the FSA project
officer during the last project meeting which is scheduled for the end of project month 6. A
final report will be completed following FSA and peer review comments on the draft report.
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8. Annexes
Annex I: Outline of the major current major technological areas or sub-fields of Synthetic biology. The analysis shown in the table is based
mainly on information published in COGEM report 2013.
Technological area or
sub-field
DNA synthesis /
Synthetic genome
What it is
The synthesis of artificial
DNA (oligos, genes or a
complete genome).
Applications
Facilitating other
applications, such
as metabolic
pathway
engineering.
Current challenges
It is not yet possible to
synthesise very long strands
of DNA in an uninterrupted
process without any errors.
For this reason shorter pieces
of DNA are produced and
then joined together. Various
techniques have been
developed to do this. The
most commonly use
techniques for DNA
synthesis are (from “small to
large”):
 chemical coupling of
nucleotides (synthesis of
Efficacy
Near future directions
The error margins
in the chemical
synthesis of DNA
are still relatively
large compared
with natural
replication145.
It is expected that the
focus of DNA
synthesis will in future
shift from the
reconstruction of copies
of existing genes (as in
genetic modification) to
the design of new genetic
circuits and functions (a
characteristic feature
of synthetic biology)146.
The error margin in the current chemical synthesis process is around 10-2 to 10-3 (or 1–10 errors per kbp) (Matzas M et al. (2010). High-fidelity gene synthesis by retrieval of
sequence verified DNA identified using high-throughput pyrosequencing. Nature Biotechnology 28 November 2010). In comparison, the error margin of natural DNA
replication in prokaryotic and eukaryotic cells lies between 10-7 and 10-8 thanks to the various proofreading and mismatch repair systems. Ma S, Saaem I, Tian J (2012). Error
correction in gene synthesis technology. Trends in Biotechnology, mart 2012 vol. 30, no 3; Schofield MJ, Hsieh P (2003). DNA mismatch repair: molecular mechanisms and
biological function. Annu. Rev. Microbiol. 57, 579-608; Li GM (2008). Mechanisms and functions of DNA mismatch repair. Cell Res. 18, 85-98; Tian J et al. (2004). Accurate
multiplex gene synthesis from programmable DN A microchips. Nature 432, 1050-1054; Carr PA et al. (2004). Protein-mediated error correction for de novo DNA synthesis.
Nucleic Acids Res. 32, e162; Hoover DM, Lubkowski J (2002). DNA Works: an automated method for designing oligonucleotides for PCR based gene synthesis. Nucleic Acids
Res. 30, e43; Xiong AS et al. (2004). A simple, rapid, high-fidelity and cost-effective PCR based two-step DNA synthesis method for long gene sequences. Nucleic Acids Res. 32,
e98.
146 Carlson R (2009). The changing economics of DNA synthesis. Nature biotechnology commentary. vol. 27:12 1091 – 1094.
145
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Technological area or
sub-field
What it is
Applications
Current challenges
Efficacy
Near future directions
Many of the
registered
components have
not been properly
characterised and
do not always
work in the ways
for which they
were designed149.
Input from
computational biology
is imperative150.
Optimised cost
production.
small fragments up to 200
bases);
 use of DNA ligases (joining
small fragments with
overlapping sequences);
 polymerisation (based on
polymerase chain reaction
(PCR)) for fragments up to
15 kilo base pairs (kbp);
 recombination (joining large
DNA fragments (from 100
kbp) or constructing a
complete genome in vitro or
in vivo (e.g. in yeast cells)).
Metabolic pathway
engineering
Also brief reference to
regulatory circuits
Inserting combinations of
genes into an organism to
introduce a new or altered
function.
Metabolic pathway
engineering takes genetic
modification a step further;
instead of inserting one or
few genes it engineers a
Fundamental
research,
production of highvalue chemicals,
biofuels,
pharmaceutical
products (e.g.
artemisinin),
bioremediation.
Optimisation of existing
components / Biobricks.
Complexity: In 2011 the
highest level of complexity of
introduced metabolic
pathways with combinations
of genes was around 10 to 15
introduced genes148.
148 Schmidt M, Pei L (2011). Synthetic Toxicology: Where Engineering Meets Biology and Toxicology. Toxicol. Sci. (2011) 120 (suppl 1): S204-S224.
149 Schmidt M, Pei L (2011). Synthetic Toxicology: Where Engineering Meets Biology and Toxicology. Toxicol. Sci. (2011) 120 (suppl 1): S204-S224; Kean S (2011). A lab of their
own. Science News Vol. 333 no. 6047 pp. 1240-1241.
150 To be suitable for standardisation and use in metabolic pathway engineering, BioBricks must possess a large number of specific properties, such as specificity, orthogonality,
sensitivity, robustness, compatibility and adjustability. In future, computer models will play an important part in the standardisation and rational design of synthetic
organisms with new functions. The results of a study to develop the first computer model of a complete bacterium were recently published. The scientists made a computer
model of the bacterium Mycoplasma genitalium which maps a considerable number of molecular interactions in the organism based on 1900 experimentally determined
parameters (COGEM report 2013),
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Technological area or
sub-field
Minimal genome
organism
Also brief reference to
orthogonal life
approaches
What it is
Applications
whole pathway into an
organism with the view to
produce a desirable output,
e.g. pharmaceutical
component, odorount, fuel,
plastic, etc.
Vehicles: Microorganisms
(bacteria, yeast), plants,
plant cell cultures.
The products of
greatest interest are
those that are
produced only in
small quantities in
their natural form
or are difficult to
process.
Regulatory circuits refers to
the engineering of novel
internal circuitry that
would enable a cell / an
organism to produce a
desired product. An
example of such circuits
and the underlying
technology is given in
Lucks et al. (2011)147.
Making a model organism
which only performs the
most essential functions
(top-down).
A minimal genome
organism is an organism
that possesses only the
most essential hereditary
Creating model
organisms,
fundamental
research.
A model organism
can function as an
optimal
chassis organism
Current challenges
Efficacy
Near future directions
Cost/benefit
analysis of the
production.
Identifying the best “model
organisms” to work with.
Standardising the minimal
genome model organisms to
employ as vehicles.
Variable efficacy
depending on the
minimal genome
organism involved.
Researchers have
succeeded in removing
about 15% to 30% of
the genome of E. coli9
and are working on a
eukaryotic model
system for a synthetic
version of the yeast S.
Lucks JB et al. (2011) Versatile RNA-sensing transcriptional regulators for engineering genetic networks. PNAS 108(21): 8617-8622. [Available online:
http://www.pnas.org/content/108/21/8617.full.pdf+html]
147
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Technological area or
sub-field
What it is
Applications
material needed to survive
in a nutrient medium. They
can only survive under
controlled conditions.
The number of necessary
genes / minimal genome
organism vary. The results
of a computer model of a
minimal cell with 241
essential genes, based on
the bacterium E. coli, were
published in 2012151.
for ‘plugging in’
natural or synthetic
genes and
metabolic
pathways
for the production
of high-value
substances153.
Potential host cells
for use as a chassis
or
production
platform include E.
coli, B. subtilis, S.
cerevisiae and
Pseudomonas
putida154.
Orthogonal life approaches
are summarised in MoeBehrens et al. (2013)152. In
brief, it refers to engineered
firewalls that would
prevent the transfer of
synthetic trains to natural
organisms/systems.
Current challenges
Efficacy
Near future directions
cerevisiae. In the
Synthetic Yeast
Genome Project 2.0,
researchers are
systematically
replacing the natural
chromosomes with
fully synthetic
genomes155.
Shuler ML, Foley P, Atlas J (2012). Modeling a minimal cell. Methods Mol. Biol. 881, 573 – 610.
Moe-Behrens et al. (2013) Preparing synthetic biology for the world. Front Microbiol. 2013: 4-5. Published online 2013 January 25. doi: 10.3389/fmicb.2013.00005 [Available
online: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3554958/]
153 Baker M (2011). Synthetic Genomes: The next step for the synthetic genome. Nature vol. 473 P403-408, 19 May 2011.
154 Royal Academy of Engineering (2009). synthetic biology: scope, applications and implications. London.
155 Posfai G et al. (2006). Emergent Properties of Reduced-Genome Escherichia coli. Science 312, 1044 (2006); Dymond JS et al. (2011). Synthetic chromosome arms function in
yeast and generate phenotypic diversity by design. Nature 477,471–476(22 September 2011); Dymond J, Boeke J (2012). The Saccharomyces cerevisiae SCRaMbLE system and
genome minimization. Bioengineered bugs 3, 168-171.
151
152
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Technological area or
sub-field
Protocell
What it is
Applications
Making a partially or entirely
synthetic cell (bottom-up).
A protocell is the simplest
artificial chemical model of
a living cell and consists of
organic and/or inorganic
elements that mimic the
function of some, but not
necessarily all, natural cell
components and molecules.
A functional definition of a
cell is that it consists of a
number of self-organising
subsystems: a metabolic
system, a form of hereditary
information for
reproduction, and a shell to
enclose and contain the
system. This enables cells to
maintain themselves, grow,
replicate and evolve156. No
protocell has yet been
developed that satisfies all
these criteria.
Fundamental
research,
developing drug
delivery systems.
Protocells are also
potential platforms
for the production
of chemical
components and
the development of
drug delivery
systems in the
medical sector157.
Current challenges
Efficacy
Near future directions
Developing a cell that is
much less complex than a
“natural” cell to facilitate its
employment for mass
production of products of
interest.
A protocell needs
to (a) have fully
functionable
membranes, (b) be
able to reproduce
(not only
replicate), (c) be
resistant to
mutations.
Progress technology to
three areas highlighted
in the column of
efficacy.
Rasmussen S, Bedau MA, Chen L et al. (2009). Protocells: Bridging nonliving and living matter. Cambridge: The MIT Press; Dzieciol AJ, Mann S (2012). Designs for life:
protocell models in the laboratory. Chem Soc Rev. 41:79-85.; Jewett MC, Forster AC (2010). Update on designing and building minimal cells. Curr Opin Biotechnol. 21: 697–
703.
157 Liu et al. (2009). Porous nanoparticle supported lipid bilayers (protocells) as delivery vehicles. J. Am. Chem. Soc. 131, 1354 – 1355; Zepik HH et al. (2008). Lipid vesicles as
membrane models for toxicological assessment of xenobiotics. Crit. Rev. Toxicol. 38, 1-11.
156
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Technological area or
sub-field
Xenobiology
What it is
Chemical synthetic biology,
or the introduction of an
alternative genetic alphabet.
The developments in
xenobiology can be divided
into various different
categories.
For example, a distinction
can be made between the
development of alternative
nucleic acids that are still
recognised by natural DNA
and RNA polymerases, and
the development of nucleic
acids that are not
compatible with the
existing natural system.
Another division can be
made according to the type
of alteration made to the
DNA:
• Modifying the structure
and composition of DNA:
creating xDNA (expanded
DNA) and yDNA (wide
DNA), for example by
adding an extra benzene
ring or fluorescent
molecule.
• Replacing the backbone:
creating xeno-nucleic acids
Applications
Fundamental
research,
production of
molecules with
specific properties.
Mainly medical
applications are
concerned.
Current challenges
XNAs are difficult to produce
in large quantities and are
usually not replicated by
polymerases.
Efficacy
Near future directions
An orthogonal system
that cannot
interfere with the
natural DNA system is
considered by some
researchers to be the
ultimate biosafety tool.
However, because so
much remains
unknown about these
new systems and their
functioning, research
will have to be done
into their
consequences for
living systems and the
environment (Is stable
replication and
functioning possible in
cells with a (partially)
orthogonal system?
What possible
interactions are there
with existing
organisms and what
will the consequences
be?).
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Technological area or
sub-field
What it is
Applications
Current challenges
Efficacy
Near future directions
(XNAs), for example glycol
nucleic acid (GNA) or these
nucleic acid (TNA).
• Expanding the codons: for
example, from four bases
(ATGC) to six bases
(ATGCPZ), and codons
consisting of four bases
instead of three bases.
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Annex II: Findings from the BBSRC/EPSRC Dialogue
(verbatim citations from Executive Summary of the report, pages 7-13)
Findings from the dialogue showed that there was conditional support for synthetic biology
- while there was great enthusiasm for possibilities of the science; there were also fears about
control; who benefits; health or environmental impacts; misuse; and how to govern the
science under uncertainty.
There was great uncertainty as to what synthetic biology would do and where it was going.
Who was driving development of synthetic biology was a big topic of debate.
Five Central questions emerged from the dialogue:

What is the purpose?

Why do you want to do it?

What are you going to gain from it?

What else is it going to do?

How do you know you are right?
A key hope was that the science could address some of the big issues facing society today,
such as global warning, serious diseases, energy problems and food security. The prospect
of being able to make progress towards these goals was a significant factor in the
acceptability of the research. (p.7)
With respect to food and crop applications specifically, the dialogue found:

Though the claims were contested, participants were initially encouraged by the
potential of synthetic biology to address issues such as food scarcity. However,
concerns arose regarding who would benefit from and own the technology.

Prominent concerns were the ability of large corporations to patent developments
and create monopolies. This could potentially maintain dependence of developing
countries on the West.

The potential impact on the surrounding environment - potentially through crosscontamination of other plants, or through pesticide resistance - was also a concern.
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
Transparency was also important in terms of food labelling so that the public could
identify food produced from synthetic biology and make choices regarding
consumption. There were concerns that ‘synthetic food’ may limit the availability of
organic or conventional crops.
Regarding regulations there was the need to open up to control to the scrutiny of others.
Ultimately, control was not just about a technical debate around risk and could not be
separated from social, economic and cultural dimensions of science. Greater thought needs
to be given to the institutional arrangements to create the conditions for synthetic biology to
be developed in useful and socially acceptable ways.
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Annex II: Agenda of the facilitated one-day meeting that was held in Fera on the 18th
November 2013 to assess current regulatory frameworks. The participants in the
meeting (shown below) represented a wide expertise. After the initial session they
were divided into two breakout groups (shown below) each of which addressed
three case studies.
13F01
10:30 – 10:45
Arrival at Fera
Coffee / tea
10:45 – 10:50
Introduction of participants – Tour de table
10:50 – 11:20
Presentation of project & results – Villie Flari
Outline of the day – anticipated outputs – V. Flari
13F01/09F01 11:20 – 12:30
13F01
Breakout groups – Case studies
12:30 – 13:15
Lunch break
13:15 – 15:00
Breakout groups – Case studies
15:00 – 15:20
Coffee break
15:20 – 16:00
Feedback to whole group (20 min per group)
16:00 – 16:45
Group discussion:
Sum up of findings and way forward
16:45
Adjourn
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Breakout groups and case studies
Breakout
groups
Group 1
Participants
Expertise
Case studies
Room
Martin Carnell
Qasim Chaudhry
GM policy department, Defra
Toxicologist and Emerging Sciences
and Technologies specialist, Fera
Which? Magazine and EFSA
Management Committee
Risk Analyst and Emerging Sciences
and Technologies specialist, Fera
Health and Safety Executive, UK
Molecular biologist, FSA advisor
Project officer, FSA
CS1.1: Coenzyme Q10 synthesised
by synthetic biology microbes.
CS 1.2: Yeast that can generate vanilla
(and potentially other) flavours in food
products in situ.
CS1.3: Enhanced levels, and
potentially a multitude of
supplements (different vitamins,
antioxidants, etc.) by a single strain
of synthetic biology probiotic
bacteria.
CS 2.1: Synthetic biology microbes
to produce new feedstock
compounds.
CS 2.2: Same as CS 1.2
CS 2.3: Non-leguminous crops able
to fix atmospheric nitrogen –
reducing the need for synthetic
fertilisers.
13F01
Sue Davies
Villie Flari (facilitator)
Michael Paton
Sandy Primrose
Jane Ince158 (11:20 – 12:30)
Group 2
James Blackburn
Christine Henry
Sarah Hugo (facilitator)
Sandy Lawrie
Claire Marris
Lei Pei
Richard Thwaites
Jane Ince (13:15 – 15:00)
GM Inspectorate, Fera
GM Inspectorate, Fera
GM Inspectorate, Fera
Regulator, FSA
Social scientist, Kings College London
Biotechnologist - Consultant,
Biofaction.com159
Molecular biologist, Fera
09F01
158
Project officer from the Food Standards Agency, UK. Additionally, Rosanna Mann and Laura Inman participated partially (during the first session of the meeting) via audio
conference.
159
http://www.biofaction.com
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