Industrial Biocatalysts for the bio­basedeconomy

Biorefinery development & Biocatalysis

Dr Pauline TeunissenRenewable Resources and BiorefineriesSeptember, 6­9, 2006York

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Genencor Division at a glance

Ø History traced to 1982

Ø May 2005: Danisco–Genencor Division:

responsible for production, innovation,

marketing and sales of industrial enzymes.

Ø 8 Bioproducts manufacturing sites; ~4 million litersof capacity

Ø Approximately 1200 Employees Worldwide

Develops and produces enzymes forindustrial applications

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Genencor & Danisco = stronger than ever

Strong financial base for investing further:

§in our innovation, applications and manufacturing capabilities

§in improved and new products

§in improving the economies of scale

We will continue to be a strong player in the industryand a reliable supplier of cost­effective performance ingredients.

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Genencor’s Products In Everyday Life

Innovative and Cost EffectiveCatalysts for Many MajorIndustrial Segments

§ Cleaning§ Textiles§ Starch Processing

§ Fuel Ethanol§ Brewing§ Leather§ Baking§ Pulp and Paper§ Food and Specialties

§ Biomass (coming soon)

Delivering on thePromise ofBiotechnology forOver a Decade!

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CellFactory

Fuels

Chemicals

Polymers

Specialties

Enzymes

Glucose

MixedSugars

Sucrose

Sugar CaneBeet

Starch Grains

Lignocellulose

The BioRefinery

Co­Products

BioRefinery

Upgraded Co­Products

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Enzymes for Fuel Ethanol Production

Enzyme§ Liquefying enzymes

§ Saccharifyingenzymes

§ Protein hydrolyzingenzymes

§ Viscosity reductionenzymes

§ Granular starchhydrolyzing enzymes

Purpose§ Thermostable alpha­

amylases for liquefaction ofstarch at high temperatures

§ Glucoamylases and blendsfor hydrolysis of starch toglucose

§ Proteases for improvedfermentation efficiency

§ Multi­component enzymesfor wheat, barley & rye

§ Alpha­amylase andglucoamylase blend forprocessing of uncookedstarch

Product nameSPEZYME® XTRASPEZYME® HPA

G­ZYME ® 480FERMENZYME ® L­400

GC 106FERMGEN

OPTIMASH ® (wheat,rye and barley)

STARGEN ® 001

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crop

GroundCorn/Starch

Saccharification

EtOHFermentation

Water / Heat

Saccharification

EtOHFermentation

Distillation

EtOH

DDG’s

Grain

Granular Starch Hydrolysis

The Opportunity:Potential to Transform the Ethanol Industry• No Cook• Fewer Process Steps• Fewer Side Reactions

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What are the potential benefits ofSTARGEN ® to the Ethanol Industry  ?

Energy saving ­ Elimination of jet cooking

Saving on the operational cost ­ Labor, time, chemicals

Value added byproduct (DDGS) ­ Higher protein content

Carbon conversion efficiency ­ Higher yield

Reduction of yeast growth inhibitors ­ High glucose, Maillard products, etc.

Reduction in by­products formation

(Reduction in osmotic stress) ­ Glycerol, organic acids, etc.

Saving on the capital cost ­ Capacity increase/new plant

Capacity increase ­ High density fermentation ­ higher alcohol yield

Process Simplification ­ Reductions in unit operations

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Enzymatic Drilling of Granular Starch

Granular Starch

Granular Starch +STARGEN ® 001,2hr

Granular Starch +STARGEN ® 001,4 hr

Granular Starch

+STARGEN, 8hr

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Comparison of different  alpha amylases withglucoamylase under yeast fermentation forEthanol

7

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0 20 40 60 80Hours

% V

/V E

toh

A.Kawachi AA

LT75

FRED

ETHYL

FA

Control

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STARGEN®001 application

§Pretreatment: Acid protease, OPTIMASH ® enzyme, acidamylase, 120 min at pH 3.6, wheat (appr. 55ºC), rye (52ºC)

§Fermentation: addition of STARGEN ® 001, 32ºC

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crop

GroundCorn/Starch

Saccharification

EtOHFermentation

Water / Heat

Distillation

EtOH

DDG’s

Grain

Waste

Starch

Oil

Fiber

Protein

The Grain Milling Process

Need: Better Enzymes andProcesses for the GrainProcessing Industry

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Technology is being brought to bear onthe issues

§ Low Energy Milling of Bio­feedstocks tosugars and other components for furtherprocessing

§ Efficient Bioconversion of Mixed Sugars toProducts

§ Efficient Utilization of Byproducts§ Optimized extraction of value§ Cost Allocation§ Bioprocess integration

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Wood chips

Switchgrass

SugarcaneCottonwoods

Corn

Paper

Sources of Cellulosic Biomass

Miscanthus

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crop

EtOH

Pre­Treatment

Saccharification

EtOHFermentation Distillation

Grain

Co­

Prod

uct

Unk

now

n Va

lue

Waste

The DOE Cellulosic Biomass Sub­Contract

Why Cellulosic Biomass:•Plentiful, and Potentially Low Cost•Could be Financially Competitive•Potential Synergies with ConventionalProcessing Technologies

The Challenge:1 billion bushel/yr Corn productionyields 25 million dry tons of Cornstover which could potentiallyproduce >2.5 billion gal EtOH peryear

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Cellulase cost was THE major bottleneck incellulosic biomass conversion

The Barrier – yr 2000

Evolution of Enzyme Cost

0

1

2

3

4

5

6

7

2000 2005

Etha

nol, 

$/ga

l

Operating Costs + DeprecEnzymesOther RawsFeedstock

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Advanced Cellulase Development Program

Project Structure and Funding§ U.S. Department of Energy (DOE) funded >$15

million over a 4­year period beginning in June 2000§ Sub­contract through the National Renewable

Energy Laboratory (NREL)§ Genencor contributed ~$5 million

Goal: Improved Low Cost Cellulase for BiomassConversion to Ethanol§ Original program targeted a 10­fold cellulase cost

reduction§ Program achieved a 30­fold reduction§ 10 – 20 cents (USD) per gallon on NREL model

substrate

Awards§ R&D Magazine, Top 100 Innovations, 2004

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Cellulase cost is no longer the major bottleneckin cellulosic biomass conversion

The Barrier Has Fallen

Evolution of Enzyme Cost

0

1

2

3

4

5

6

7

2000 2005

Eth

anol

, $/g

al Operating Costs + DeprecEnzymesOther RawsFeedstock

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Technical Strategy

§ Engineer & recruit improved cellulase components§ Protein Engineering:  Improved thermostability in CBH1 &

CBH2§ Novel Cellulolytic Activities A. cellulolyticus E1

§ Develop an enhanced T. reesei production organism toproduce all the key activities in one host§ Strain construction§ Fermentation & product performance screening

§ Improve the Manufacturing Process to minimizeproduction cost§ Fermentation & Strain improvement including genomics,

mutagenesis and screening

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0

50

100

150

200

250

300

350

400

450

500

0 10 20 30 40 50 60

Program Months

2.3X

11X20X 30X

Combined Enzyme step cost drop 30X

$/ga

l EtO

H(N

RE

L M

etric

)

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Overall Cellulase Cost Reduction

~30XTotal

~4 X

Improved enzyme performance­ 41G enzyme mix­ Recruited cellulase E1­ 56 oC vs 38 oC conversion

~8X

Improved protein production­ Whole cell product (i.e., no recovery)­ Media improvements­ Carbon source­ Strain productivity (41G)­ High cell density fermentation

ImprovementFactorImprovement

Audited by

NREL

Validated by

NREL

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Enzyme cocktail tuned tofeedstock & pretreatment

Biomass Value Chain Integration

FEEDSTOCK

PRETREATMENT

ENZYMEHYDROLYSIS

FERMENTATION

END PRODUCTS

Corn stover,bagasse, pulp, etc.

Mechanical orthermo chemical

Complex sugars tosimpler sugars

Yeast to ethanol (C­6) Need a C­5 plus C­6 bug

Ethanol and other biochemicals

FEEDSTOCK PRETREATMENT FERMENTATION END PRODUCT

CurrentGCOR role

ENZYMEHYDROLYSIS

Process choice needed

No infrastructure for someNo collection implements

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Further Issues… .

Biorefinery Issues Facing Genencor and Others§ Supply Chain

§ What will drive raw material availability?§ What incentives or guarantees will be required?§ What is “value” of each player’s contribution§ Whole broth enzyme formulation

§ IP protection§ Co­location and/or single purpose dedicated facility

§ Demand Chain§ What in addition to ethanol will provide the best

value to investors?§ What will it take for them to be accepted in the

market?§ Investment

§ What is best mechanism to allow regional/localinvestors to participate?

§ What is the role of investment types (angel,traditional, etc.)

§ Derisking funds – federal and state (pioneers face hugerisk)

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crop

GroundCorn/Starch

Saccharification

EtOHFermentation

Water / Heat

Saccharification

EtOHFermentation

Distillation

EtOH

DDG’s

Pre­Treatment

Saccharification

EtOHFermentation Distillation

Grain

Co­

Prod

uct

Unk

now

n Va

lue

Waste

Starch

Oil

Fiber

Protein

The BioRefinery Front­End

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Conclusions

§ Technologies are rapidly advancing.

§ Biomass conversion technology continues toprogress. Significant challenges remain.

§ Starch processing technology is rapidlyevolving.

§ Cellulosic & starch technologies & processescould very well merge in synergistic ways.

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Biotechnology offers Sustainable Solutions toGlobal Challenges

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Genencor’s vision

…  a leading innovator of products for

the bio­based economy.

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Genencor’s vision

Ø We envision a future where biotechnology fulfills manyunmet needs..Ø In development and deployment of industrial enzymesØ In the production of fuels, chemicals & materials

Ø We envision a future where biotechnology helps createsustainable industrial activities

Ø We envision a future where biorefineries take theirplace alongside oil refineries

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