Emerging Business Area White Biotechnology Industrial Biotechnology Towards Next Generation Biorefineries

Marcel Wubbolts DSM White Biotechnology b.v. Delft, The Netherlands

DSM White Biotechnology

Chemelot Colloquium, October 23th, Geleen

DSM Innovation Center, HQ in Urmond

Chief Innovation Officer
Staff

P&T Business Incubator Licensing

EBA Biomedical Materials Urmond, NL EBA Personalized Nutrition Parsippany, US (Delft, NL) EBA Specialty Packaging Urmond, NL

Corporate Technology

Venturing

EBA White Biotechnology Delft, NL

DSM White Biotechnology

Location of DSM White Biotechnology b.v.

DSM White Biotechnology b.v., HQ in Delft (NL) Part of DSM Innovation Center b.v. Urmond (NL) White Biotechnology Beijing (CHN)

White Biotechnology Geneseo, IL (US)

White Biotechnology Freemont, CA (US)

DSM White Biotechnology

Recent Press Message - 211008

DSM White Biotechnology

Location of DSM White Biotechnology b.v.

DSM White Biotechnology b.v., HQ in Delft (NL) Part of DSM Innovation Center b.v. Urmond (NL) White Biotechnology Beijing (CHN)

White Biotechnology Geneseo, IL (US)

White Biotechnology Freemont, CA (US)

White Biotechnology Belvidere, NY (US)

DSM White Biotechnology

Colours in Biotechnology

White Biotechnology - Industrial Biotechnology Red Biotechnology - Healthcare Biotechnology Green Biotechnology Plant biotechnology

DSM White Biotechnology

What about White Biotechnology ?

Industrial or White Biotechnology is an emerging field within modern biotechnology that serves industry. It uses living cells like moulds, yeasts or bacteria, as well as enzymes to produce goods and services

source: DSM White Biotechnology

Living Cells & Enzymes

Cell (mini-factory)

Enzyme (catalyst)

Enzyme (structure can be designed and changed by laboratory evolution) DSM White Biotechnology

Industrial (White) Biotechnology

Starting Material

Process

Products

starch

BioBased Chemicals BioFuels

biomass

BioProducts BioMaterials

other (renewable) materials DSM White Biotechnology

Why is White Biotechnology so Important to DSM?

White Biotechnology, in contrast to Petrochemistry, makes use of a renewable resource: sunlight via agricultural feedstocks The use of crops / crop residues as feedstock is an alternative to crude oil (volatile prices) and leads lower CO2 emissions.

White Biotechnology is a powerful source of innovation for new products and processes and is a barrier to entry for competitors. DSM has strong capabilities in White Biotechnology via its Life Sciences technology base. DSM has leading competences in Chemistry and Materials Sciences that become increasingly integrated with Life Sciences.

DSM White Biotechnology

Crude Oil Prices (October 2008)

DSM White Biotechnology

Industrial Biotechnology Penetration

Source: Ceasar, Industrial Biotechnology, March 2008

DSM White Biotechnology

Industrial Biotechnology Segments More than Biofuels

Source: Ceasar, Industrial Biotechnology, March 2008

DSM White Biotechnology

Innovation is our SportTM

DSM White Biotechnology

DSM: Ability to change

100 years of successful transformations Evolution
Bioterials / Biologics Life Science Products Performance Materials Petrochemicals Fertilizers Coal

1902

1930

1950

1970

1990

2000

2010 Technological competences

Mechanical engineering Chemical engineering Polymer technology Material science Fine chemicals Biotechnology

DSM White Biotechnology

DSM: Ability to change

100 years of successful transformations Evolution
Bioterials / Biologics Life Science Products Performance Materials Petrochemicals Fertilizers Coal

1902

1930

1950

1970

1990

2000

2010 Technological competences

Mechanical engineering Chemical engineering Polymer technology Material science Fine chemicals

Classical Biotechnology

Modern Biotechnology

DSM White Biotechnology

DSM has a long history in Biotechnology

Classical Biotechnology (>1870s)

Yeast, Ethanol, Yeast extracts, Vitamins, Penicillin, Butanol (ABE), Enzymes, Citric acid

Modern Biotechnology (>1980s)

Recombinant enzymes (e.g. Chymosin, Phytase) Metabolic engineering (e.g. Vitamin B2, Cephalexin) Biocatalysis (e.g. pharmaceutical intermediates) Cell culture (e.g. Per C6TM human cell line)

White Biotechnology (>2005) 1. Enzymes & Yeast for next generation biorefineries 2. Renewable processes for chemicals, materials and biofuels.

DSM White Biotechnology

DSM Nutrition Biotechnology Process: Vitamin B2

Fermentation of a Natural Compound
Sugar
Metabolic Engineering

Bacillus subtilis

12 raw materials replaced by sugar 7 solvents replaced by water waste reduction of over 75%
20 m

cost reduction of over 20%

Vitamin B2

environmental impact 40% lower

DSM White Biotechnology

DSM Pharma Biotechnology Process: Cephalexin antibiotic

Fermentation of a Non-Natural Compound

Old Route Sugar

Traditional Fermentation

New Route

13 chemical steps Metabolic Engineering replaced by: 1 fermentation + 2 enzymatic steps

2 enzymatic steps

Sugar

Penicillium chrysogenum

13 chemical steps

65% less energy and materials 50% lower cost

DSM White Biotechnology

Vision for EBA White Biotechnology

DSM to build a leadership position in selected White Biotechnology Fields by enhancing existing internal capabilities through key external partnerships: BioBased Chemicals and Advanced BioFuels in partnership with upstream and downstream players Provider of Yeast and Enzymes for cellulosic ethanol production

DSM White Biotechnology

White Biotechnology and Tools for Biofuels

Provider of Yeast and Enzymes for cellulosic ethanol production

DSM White Biotechnology

Raw material transition

Coal
Non-renewable

Oil

Biomass
Renewable

DSM White Biotechnology

GHG Emissions (CO2 eq.)

Source: http://ngm.nationalgeographic.com/2007/10/biofuels/biofuels-interactive

DSM White Biotechnology

Amount of Crop Residues Available: Origin Corrected for need to prevent erosion / fertilizer

Source: LMC Starch & Fermentation Analysis, June 2008

DSM White Biotechnology

Industrial Biotechnology Trend towards Cellulosics

Second generation biorefineries (based on waste biomass, 2nd generation) for chemicals and fuels production are crucial to cope with crude oil shortages and avoid (perceived) food competition. Intelligent use of biomass is needed to address issues:
Land use Food versus Fuel Water consumption Sustainability certification for Biomass Higher efficiency in agricultural productions (globally) Efficient use of biomass Innovations in cellulosics processing, hydrolysis and fermentation New (and existing) biobased products

Innovations needed to make this happen

DSM White Biotechnology

Oil-Refineries vs Bio-Refineries
Step Up in Efficiency Required
Oil-refinery:
Crude oil (finite) as input Established technology Very efficient use of raws

Bio-refinery (1st generation):

Starch / Sugar (renewable) as input Technology established Efficient use of feedstock

Bio-refinery (2nd generation):

Biomass (renewable) as input Logistics? Technology very much in development (Enzymes & Fermentation Organisms) What do we do with byproducts?

DSM White Biotechnology

Current (1st) and Future (2nd generation) Feedstock Use Corn as an Example
Current (1st)
Starch Enzymes Starch C6 Sugar C6 Yeast

BioEthanol

Corn

Corn Kernels

BioSuccinic Residue
Biomass

Future (2nd)
C5 Yeast C5 Yeast Cellulose Enzymes Hydrolysate C5 sugars C6 sugar Bacteria

BioButanol

Other

Pretreatment

DSM White Biotechnology

Pre-treatment
Room for innovation

Popular method: mild acid pretreatment (0,5 vol. % H2SO4, 180-190C)

Short residence time Cellulose intact, hemicellulose hydrolysis Some inhibitor formation (acetic acid, furfural, HMF) Waste production (CaSO4)

Source: Michael R. Ladisch, Nathan Mosier, Gary Welch, Bruce Dien, Andy Aden, Phil Shane, Purdue University

DSM White Biotechnology

Enzymatic hydrolysis (30 L)

high solids required: operational challenges

DSM White Biotechnology

Cellulosic Material as Feedstock Innovations needed for 2nd generation to happen!

C5 Yeast

Cellulose Enzymes

It is not that easy at all and requires substantial innovations

DSM White Biotechnology

DSM Enzymes for Wave 2 Feedstocks

Leveraging DSMs Enzyme Technology
DSM has large collection of hydrolases (cellulase, hemicellulase, pectinase) and fermentation processes developed for various business segments Several existing DSM in house enzyme products of value for cellulose processing US DoE awarded R&D grant to DSM White Biotechnology for further development of enzymes for second generation feedstocks
A. niger sequence: Pel et al. (2007) Nature Biotechnology 25, 221 P. chrysogenum sequence: van den Berg et al. (2008) Nature Biotech. 26, 1161

Cellulose Enzymes

DSM White Biotechnology

DSM White Biotechnology Press Cellulose processing enzymes: DSM and Abengoa New Energy (DoE grant awarded tot DSM, 27 February 2008)

DSM White Biotechnology

How to develop the right enzymes?

Key to success: Diversity
- Large libraries of (bio)catalysts required (in house / partners) - Access to genomes and meta-genomes (incl. bioinformatics) - Directed evolution to optimize biocatalysts

Screening power
- Automation and miniaturization - High through-put screening - Fast, reliable and generic analysis (NMR, LC-MS/MS)

Process optimization & scale-up

- PluGbugTM - High through-put experimentation (Table Top Biorefinery) - Engineering & modeling

DSM White Biotechnology

Generic Analysis: Flow Injection NMR

Joint development of DSM-Gist and Bruker: BEST-NMR Bruker Effcient Sample Transfer (based on Gilson XL215) Now commercially available

DSM White Biotechnology

Screening & Strain Development

DSM White Biotechnology

Table Top Biorefinery

Pretreatment

Ethanol fermentation (AFM)

DSM White Biotechnology

Corn Fiber Pretreatment

Original 90% DM

Milled 90% DM

Destarched 25% DM

Cooked & Saccharified 12.5% DM

Pretreatment & enzyme treatment result: strongly reduced viscosity

DSM White Biotechnology

Eschweiler 10L Fermentors in Delft

DSM White Biotechnology

4m3 - 100m3 Fermentors

DSM White Biotechnology

Proprietary DSM Enzymes applied in Lignocellulose Hydrolysis

Dilute acid pretreated lignocellulosic feedstock converted at 10% dry matter DSM cellulases added and incubated for 16 hrs at 65 C, pH 5.0 to reduce viscosity and produce monomeric sugars for fermentation DSM C6-Yeast in Simultaneous Saccharification and Fermentation for 150 hrs at 33 C Complete cellulose conversion in Simultaneous Saccharification and Fermentation (SSF) DSM White Biotechnology

(Ligno) Cellulosic Feedstock Composition Corn as an Example

C5 C6

C6

Cellulose 20-45% (C6) Hemicellulose 20-55% (C5)

DSM White Biotechnology

C5

Fermentation Organisms for 2nd Generation Feedstocks

Leveraging DSMs Bakers Yeast Technology Position Traditional Ethanol Yeast
Long history of safe industrial use Fermentation and strain development tools well developed Robust organism Used at low pH, which prevents infections (non-sterile) Facultative anaerobe (no aeration needed, lower CAPEX, energy input low) First eukaryotic genome sequenced (1998) >40 mio tpa fuel ethanol applications for > 30 years Efficient fermentation of C6 sugars (1st generation)

However, traditional Yeast only uses C6 sugars and it does not efficiently produce BioEthanol on cellulose feedstock (C5 sugars)
C5 Yeast

DSM C5 Ethanol Yeast

DSM is developing Yeast that consumes cellulose hydrolysates (C5 Yeast), suited for 2nd generation biorefineries

DSM White Biotechnology

Example of Metabolic Engineering of Utilization of C5 Substrates in Saccharomyces cerevisiae

DSM White Biotechnology

White Biotechnology and BioBased Chemicals BioSuccinic Acid

DSM White Biotechnology

Environmental Advantages of Biobased Chemicals

Green House Gas (GHG) Emission Reductions of Bio-succinic Acid
Succinic acid from renewable feedstocks vs petrochemical feedstocks
GHG emissions (t CO2,eq/t product) 7 6 5 -50% 4 3 2 1 0
Pe tro ch em ic al C or n -s ta rc h ne ca
GHG emissions (t CO 2,eq/t product) 7 6 5 4

-65%

-65%
3 2 1 0
Pe tro ch em ic

-80%

75%

Today's technology (2005) Today Today Today

Future

Future technology (2030) Future Future

Source: Patel et al. (2006). BREW Report; and Hermann, Blok and Patel (2007). Environ. Sci. Technol. 41, 7915-7921.

DSM White Biotechnology

o (2 n rn d G sto en ve .) r

rc h

al

Su

Su ga rc an

ga r

or n

-s ta

Future

DSM White Biotechnology Press Biosuccinic acid: DSM and Roquette (18th January, 2008; BioHUB program)

DSM White Biotechnology

DSM-Roquette Partnership for Bio-Succinic Acid

Bio-Succinic Acid Intermediate for BioBased Materials
Biorenewable Polymers (e.g. PBS) Renewable Feedstock Bio-SA Fermentation Existing large-volume products like butanediol (BDO) Bio-SA derivatives (e.g. solvents, various esters)
RQ-DSM Alliance

RQ

DSM & downstream partners

DSM White Biotechnology

White Biotechnology and BioBased Chemicals BioButanol

Biobased Chemical and a Potential Advanced Biofuel

DSM White Biotechnology

Global Butanol Demand: Approximately 3000 kt - $4 billion market

GROWTH 3%- 4% / yr. +75 +120 +115 +150
Central and South. America only 80 kt Supply gap in China and Europe despite planned expansions CHN: largest growth & planned capacity expansions

4500

3940
4000
ROW 420

3500 V o lum e (k t) 3000 2500 2000 1500 1000 500 0

3060
ROW 345 Europe 600 Asia 630

Europe 720

Asia 745

USA 940 USA 790 China 1115 China 695

+420 +880 KT

2007

2015

Quality will be important for chemical market penetration

Sources: SRI/NEXANT & DSM analysis

DSM White Biotechnology

White Biotechnology and Materials

BioBased Chemicals and Advanced BioFuels in partnership with upstream and downstream players

DSM White Biotechnology

A New Value Chain is Emerging

Feedstock Provision

Feedstock Processing

Primary Conversion

Secondary Conversion

BioCompounds & Formulation

Converters

Farming Storage Distribution

Grain seed milling

Biomass pretreatment - enzymatic conversion

Fermentation

Polymerization

Compounding

Moulding Film extrusion

Chemical conversion Chemical Synthesis Formulation Enzymatic conversion

Reactive Extrusion

Food & Feed

Bio Medical

Coatings

Automotive

Life Sciences
Pharma Personal Care Electrical

Material Sciences

DSM has multiple capabilities that extend across several stages in the valuechain that extend into multiple product and market combinations.

DSM White Biotechnology

Conclusions
Bioprocess economics and environmental advantages such as fossil energy savings and greenhouse gas reductions make the production of biobased chemicals, biofuels and materials using White Biotechnology desirable Life Sciences and Materials Sciences are becoming increasingly integrated, opening new opportunities for Industrial Biotechnology. At DSM most of the required competences are available in house. Additional development and innovations at multiple positions in the value chain, including lignocellulosic feedstock (2nd generation) processing, are needed to fully exploit the potential of Industrial Biotechnology in chemicals, biofuels and materials applications

DSM White Biotechnology

DSM White Biotechnology