October 2010
Development of Enzymes for Biomass
Degradation and other Bioenergy
Activities at Novozymes
JOHAN
JOHAN MOGENSEN,
MOGENSEN, DEPARTMENT
DEPARTMENT MANAGER
MANAGER
BIOFUEL
BIOFUEL ADU-R&D,
ADU-R&D, NOVOZYMES-DK
NOVOZYMES-DK
BioEnergy R&D activities within 4 areas
•
Cellulosic bioethanol
• Starch based bioethanol
• BioDiesel
• Biogas
2
Novozymes and BioEnergy
Largest supplier of enzymes to the “starch” fuel
ethanol industry
Approx. 18% of total sales in 2009
Mainly a US business
New processes reduce energy need and
improve yields from corn-based production
Collaborations in cellulosic ethanol with industry
leaders
Enzymes for commercial production of cellulosic
ethanol ready by 2010
Global biofuel production has grown rapidly and will
continue to grow – great potential for enzyme sales
worldwide
1G ethanol
2G ethanol
Biodiesel
350
300
250
BLY
200
150
100
50
0
2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019 2020 2021 2022
Sources: VOPAK, Emerging Markets Online, F.O. Licht, Novozymes estimates
Conventional Starch Biofuel vs. Advanced
Biofuels From Biomass
Readily available starch is hydrolyzed
by enzymes into fermentable sugars
Enzyme
process
Fermentation
Pretreatment
process
Cellulose
Bioethanol
Enzyme
process
Enzymes break down the sturdy cellulosic material into cellulose, hemicellulose,
and lignin which then is hydrolized to sugars. Enzyme need is significantly
higher than the conventional process.
Cellulosic Ethanol
6
Applied discovery reduce total process costs
Novozymes’ core
• Highly interrelated technologies
• Existing R&D tools and experience utilizing
enzyme technology and application know-how
Cellulose Hydrolytic System
Synergy
CBH II
EG III
EG I, II
CBH I
Approaches to making enzymatic hydrolysis viable for
B2E – explored by Novozymes
Improving enzyme performance
• Additional activities to existing efficient cellulytic complex
Improving process compatibility with enzyme hydrolysis
• Separate hydrolysis and co-fermentation of C5&C6 sugars
Improving enzyme production cost
• Optimizing expression yields
Advantages and disadvantages of T. reesei
Advantages
• Exceptionally high secreted protein expression potential
• Base cellulase system has very good specific activity compared to other
cellulolytic fungi
• Genome sequenced and well annotated
Disadvantages
• Most secreted proteins not particularly thermostable
• Native ß-glucosidase and hemicellulase secretion insufficient for most
substrates
• Native GH61 proteins not highly expressed and not particularly active
• Other organisms produce enzymes that are individually superior
ß-glucosidase screening
B
ce ase 1X
llu
las 2X
e
Cellulose hydrolysis
NREL PCS, 50 °C, pH 5.0, 120 hr
ßG performance in high liquor and glucose
5% IS + liquor equivalent of 20% TS + glucose at 40 g/L
NREL PCS + high liquor + high glucose, 72 hr
14
Glucose release (mg/mL)
12
10
8
6
4
2
0
0.0%
1.0%
2.0%
3.0%
% BG Addition
4.0%
5.0%
GH7 CBH I in high temperature mix
Glucan conversion, %
Top replacement candidate
T. reesei CBH I
No CBH I
(3 mg/g)
Temperature, °C
CBH I’s (1.05 mg/g cellulose) were added to mono-component cellulase mix lacking
CBH I (1.95 mg/g).
NREL PCS hydrolysis, 72 hr.
Expression of heterologous CBH I in T. reesei cbh1
deletant strain
Top candidate CBH I
“If it doesn’t express, it doesn’t exist”
CTec 2009 vs. CTec2: enzyme dose reduction on a
range of commercially relevant substrates
Feedstock
Feedstock data
Pretreatment
Method
Washed/
Unwashed?
Measurement
% glucan
conversion
Corn Stover
Acid
unwashed
80%
Corn Fiber
Acid
unwashed
70%
80%
70%
72%
Corn Cob
Acid
unwashed
Wheat straw
Steam
washed
80%
Corn Cob
Steam
unwashed
80%
Corn Stover
Steam
washed
80%
Hardwood
Organosolv
washed
70%
Sugar Cane
Acid/Alkaline
washed
80%
Fold
Reduction
1.3
1.4
1.3
2.0
2.6
2.5
2.0
1.5
1.8
1.4
How much better will the next generation be?
The strains co-expressing multiple improved proteins are under
construction
We anticipate substantial reductions in protein loading, but they
are likely to be substrate and pretreatment-dependent
Optimum hydrolysis temperature will be higher, presumably
resulting in lower contamination probability and perhaps lower
substrate viscosity
STARCH BASED
BIOETHANOL
R&D activities
17
Ethanol Yield Potential Definition
In the current conventional fuel
process there is an additional
theoretical yield of 7 % which is
not turned into ethanol.
The average plant conversion of
starch to ethanol is 87 %
Starch to ethanol
87 %
Approx 6-7 % of the glucose
pool is lost to yeast growth and
glycerol production
Yield enhancing concepts mainly
access the remaining ~7% of starch
6-7 %
6-7 %
BIODIESEL
R&D activities
19
Global biofuel production has grown rapidly
and will continue to grow
(Ethanol volume projection are 4*biodiesel)
1G ethanol
2G ethanol
Biodiesel
350
300
20 %
BLY
250
200
150
80 %
100
50
0
2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019 2020 2021 2022
Sources: VOPAK, Emerging Markets Online, F.O. Licht, Novozymes estimates
Key advantages with enzymatic biodiesel
•
•
•
•
•
Compatibility with variations in the quality of the raw material
Reduced energy consumption and waste water volumes
No strong acids and bases
Allow for ethyl esters (FAEE)
Higher quality of glycerol
We believe enzymatic biodiesel can be made costeffective on an industrial scale
1. Low-cost immobilized enzymes
2. Process development (how should the enzymes be applied)
Conclusions
• In spec FAEE can be produced enzymatically.
• The economy suggests the enzymatic process can be cost
effective.
• Enzyme technology can improve the environmental sustainability
of the biodiesel production and the use of the fuel.
Biogas at Novozymes
Focus in Novozymes
Enzymes and microorganisms
CH4
Manure
CH4
Sugar Cane
Bagasse
Pretreatment
Energy Crops
Anaerobic digestion
Secondary digester
Novozymes technology for biogas not
identified/confirmed yet but first
leads and strong synergies/overlap to
other industries (eg. Bioenergy)
27/
10/
201
0
Waste water
Thank you
Join the dialog…
www.bioenergy.novozymes.com