"From the Investigation of marine model species to biotechnological developments, a knowledge‐
based approach"
Catherine Boyen
OECD Global Forum on Biotechnology: Marine Biotechnology Enabling Solutions for Ocean Productivity and Sustainability (Vancouver, Canada, 30‐31 May 2012)
Outline
Marine Diversity, Evolution and Environment
Marine Models and Biotech :
‐ Marine Macroalgae
Polysaccharides/Enzymes
From Academia to Biotech Development
4 projects funded by the French government & CE
‐Bioresources and biotech
IDEALG
OCEANOMICS
‐Infrastructures
EMBRC
GREENSTARS
Marine Diversity and Evolution
Oceans ….. the craddle of life
Prokaryotic life originated in the oceans about 3.6 billions years ago Eukaryotic life: between 3 and 3.1 billions years ago
Land became colonized by fungi about 1 billion years ago and green plants only 700 million years ago. Eukaryotic tree
[Cock and Coelho, 2011, J Exp Bot]
Marine diversity and environment
COOH
Habitat Diversity
Salinity
Pressure
Light
Temperature
Tides
Abiotic stress
Signalling/Communication
Secondary metabolites
Biotic stress
Grazing
Biofilm
Larval settlement
POLYSACCHARIDES from MARINE ALGAE From Basic Research to new developments in Biotechnology
From the study of cell wall to the discovery of new active molecules
FROM ACADEMIA to the INDUSTRY
Diversity of Polysaccharides in Plants/Seaweeds
Land plants
Green algae
Red algae
Cellulose
Cellulose
Mannan
Xylan
Brown algae
Neutral polysaccharides
Cell wall
skeleton
Cellulose
Cell wall matrix
(hemicellulose)
Xyloglucanes
Mannane
‐(1,3)‐(1,4)‐
glucane
‐(1,3)‐glucane
Carbon storage
Starch
Xyloglucanes
Mannane
Glucuronane
-(1,3)-glucane
Starch
Glucomannane
‐(1,3)‐(1,4)‐
glucane
‐(1,3)‐(1,4) xylane
Cellulose
-(1,3)-glucane
Floridean Starch
Laminarin

Alginates
Anionic polysaccharides (cell wall matrix)
Carboxylic
Sulfated
Pectins

Ulvans
Ulvans OSO3-
Agars
OSO3OSO3Carrageenans
Sulfated fucans
Porphyrans
Diversity of polysaccharide‐degrading hydrolases
Species
Enzymes
End products
Pseudoalteromonas carrageenovora
Kappa-carrageenase
Lambda-carrageenase
Kappa-neocarrabiose
Lambda-neocarrabiose
Alteromonas atlantica
Beta-agarase I
Beta-agarase II
Neoagarotetraose
Neoagarobiose
Alteromonas fortis
Iota-carrageenase
Iota-neocarratetraose
Alteromonas beaufortensis
Beta-agarase
Neoagarotetraose
Alteromonas agarilytica
Alpha-agarase
Agarotetraose
Zobellia galactanovorans
Kappa-neocarratetraose
Iota-neocarratetraose
Neoagarotetraose
Neoagarotetraose
Marineflexile funavivorans
Kappa-carrageenase
Iota-carrageenase
Beta-agarase A
Beta-agarase B
Porphyranase
Fucanase
Pseudomonas alginovora
Alginate-lyase
MM MG
DP 4
POLYSACCHARIDES from MARINE ALGAE + Specific enzymes (hydrolases) from Marine Bacteria
A large diversity of Oligosaccharides New Biological activities
Iodus 40®,
First defence stimulating product for wheat
On the market since 2003
Active molecule = LAMINARINE (oligosaccharide from Laminaria digitata)
CH 2OH
CH 2OH
O
O
OH
OH
OH
OH
ß-1,3 glucane
Control
:
Induced :
New Enzyme discovery in the Pre‐genomic Era
Identification of polysaccharide hydrolases
Given the scarcity of gene sequences with a validated activity, classical purification still
remains the most direct way for expanding our
catalogue of enzymes acting on algal polysaccharides.
E., g., isolation of the l‐ carrageenase from
Pseudoalteromonas carrageenovora
0,16
0,14
1
2
3
4
5
140
120
66
45
0,12
A220n m
Incubati on
ti me
200
116
97
100
0,10
80
0,08
60
0,06
carrageenase activity (U)
Protein size (kDa)
0’ 5’ 10’ 20’ 30’ 45’ 60’ 2h 4h 6h 8h 12h 24h 48h 96hT96h
DP3
Neo-carrahexaose
40
0,04
DP2
Neo-carratetraose
20
0,02
0,00
0
15
16
17
18
19
Elution volume (ml)
20
21
A
B
Guibet, Colin et al.,
Biochem. J., 2007,
…With genomic resources
Ectocarpus siliculosus genome 190
Mpb, Cock et al (2010) Nature
Zobellia galactanivorans: a marine flavobacterium
isolated from the red alga Delesseria sanguinea;
Barbeyron et al. (2001) IJSEM
Census of Ectocarpus CAZymes
GH1
GH2
GH3
GH5
GH10
GH16
GH17
GH30
3
2
1
2
1
6
1
1
GH31
1
GH36
GH37
GH38
GH47
GH63
GH81
GH85
GH88
GH95
2
1
1
5
1
5
1
6
1
GT1
GT2
GT4
GT7
GT8
GT10
GT13
GT14
GT15
1
11
13
1
3
1
2
5
1
GT20
GT22
GT23
GT24
GT25
GT33
GT34
GT41
GT47
6
3
2
1
3
1
1
3
8
GT48
GT49
GT50
GT54
GT57
GT58
GT59
GT60
GT64
1
1
3
2
3
3
1
1
2
GT65
GT66
GT74
GT76
GT77
1
1
1
1
1
Census of the Zobellia CAZymes
• 114 glycoside hydrolases, 12 Polysaccharide lyases
and 17 Carbohydrate Esterases !!!
♦ several large multigenic families
GH16 (agarases, -carrageenases, laminarinases)
GH29 (L-fucosidase), GH43 (xylosidases, arabinases)
GH13 and GH97 (starch degradation), PL1 and PL10 (alginate lyases)…
GH = Glycoside hydrolases 41
• 72 sulfatases ! (1,5 % of the genome)
GT = Glycosyltransferases 88
Michel G. et al New Phytol, 2011
NEW ENZYMES + POLYSACCHARIDES + OMICS
NEW OLIGO‐SACCHARIDES
NEW BIOLOGICAL ACTIVITY
COSMETIC
NUTRACEUTIC
ANIMAL HEALTH
“Investments for the future” program
35 Billions €
For supporting Research Infrastructures and Innovation
 9 large calls (2 rounds: 2010 & 2011)
 Funding for 8‐10 years
3‐ HEALTH and BIOTECHNOLOGY
‐National Infrastructure in Biology and Health ( ESFRI)
‐Biotechnology and Bioresources
4 ‐ENERGY
‐Institute of Excellence in Decarbonated Energy
2011 – 2020 www.idealg.ueb.eu
Scientific coordinator Dr Philippe Potin (CNRS, Roscoff)
Make the best of Omics research applied to seaweed and associated bacteria to generate new genetic tools and culture methods for improving algal bio‐resources and the domestication of local seaweed species and to develop blue and white technologies
Include environmental impacts studies as well as economic, social and ethical issues in order to guarantee a sustainable development of the algae sector
12 academic partners
5 private partners
1 technical center Aim : A chain of transfer of knowledge
Seaweedomics
towards
Systems biology
Domestication
Biotechnology
Sustainable
Chemistry
Develop basic research on brown, red and green seaweeds toward
domestication of local crops and improvement of seaweeds uses in biotechnology and blue‐green chemistry
G greenstars
S
Micro-algae and by products
a large network of partners
Innovation for industrial biorefinery of microalgae
« Investissements d’Avenir » Institutes of Excellence on Decarbonated Energy)
The core idea of GreenStars
To build the missing link on ‐algae between
academic research and industry
From physiology of microalgae to environmental biorefinery
Multiple targeted markets (animal food, cosmetic, green chemistry, energ
GreenStars, several exploration platforms
Strain collection
Heliobiotec
TIP
Lagrangian simulator
Algotron
Aquaculture
GreenStars, applications are everywhere !
3 years
5 years
10 years
7 years
Time of access to market
Energy
Green Chemistry
Cosmetic
Nutrition
High volume markets
Lower added value molecules
Aquaculture
Challenges:
‐ adapted species
‐ recovery, extraction
‐ low production costs
Biogas
Animal feed
Low volume markets
High value molécules
« European Marine Biological Resource Centre » (EMBRC)
on the road map of « European Strategy Forum for Research Infrastructures » (ESFRI) since 2008. A preparative project since 2010
http://www.embrc.eu/
‐ EMBRC : a distributed Research Infrastructure of state‐of‐the‐art research and training facilities at leading marine research stations across Europe.
‐ EMBRC will provide End‐Users from Small and Medium Enterprises (SMEs), Academia and Industry with access to marine biodiversity, its associated meta‐
data and extractable products.
SERVICES: ACCESS TO:
Pho
to : L. LEV
EQ
UE
Ecosystems
Marine Resources
Technological
PF
Biobanks
In situ experimentation
Hosting
facilities
Bergen
*
** *
*
*
*
*
*
*
*
Oban
St. Andrews
Plymouth
Tjärno‐Kristineberg
Bremerhaven
Roscoff Villefranche
Banyuls
Faro
Naples
Crete
OCEANOMICS
2012‐2020
WOrld oCEAN biOressources, biotechnologies, and Earth‐systeM servICeS
Biotechnology and Bioresources for Valorization of Marine Planktonic Ecosystems
2009
2010
TARA‐OCEANS
10 Million €
2011
2012
2013
2014
2015
2016
2017
2018
2019
2020
OCEANOMICS
7 Million €
BIO‐MONITORING
BIO‐PROSPECTING
High‐Throughput Screening
OCEANOMICS WORK PLAN
160 Stations  40 093 biological samples (various depths and size fractions)
Total of 426,885,078 ILLUMINA sequences, corresponding to 9,074,257 distinct sequences
each sequence is present 47 times in average
THANK YOU