A New Biorefinery Platform Intermediate

Principal Investigator: Doug CameronProject Managers: Jim Millis, Paris Tsobanakis

2005 OBP Bi-Annual Peer ReviewIntegrated Biorefinery Platform

November 16, 2005

Overview

Scope of Work Plan

• Cargill, in partnership with Codexis, is developing a fermentation route to a novel platform chemical, 3-hydroxypropionic acid (3-HP)

• We are also collaborating with PNNL to develop novel catalytic process for the conversion of 3-HP to acrylic acid and acrylamide

Glucose 3-HPAcrylic AcidAcrylamide

Microbialfermentation

Chemical catalysis

Overview

• Award was for $6.09 million in matching DOE funds for a 3 year program (total $12.2 MM), representing a 50/50 cost share

• Biotech/Codexis work started in May 2003; Chemical conversion/ PNNL work started in December, 2003

• The pace of the program was slowed after one year as commercial activities lagged technology development. ‒ $0.60 MM received in FY03 (May to September)

‒ $1.66 MM received in FY04

‒ $0.77 MM received in FY05

‒ Request for FY06 and beyond to be submitted by end of November

‒ 50% complete

‒ Completion of the program will require a one year extension

Overview

• The program is currently in Development Phase

• Barriers addressed

3-HP is a novel platform chemical (identified as a Top Ten product) with the potential to impact both large, commodity markets and new, emerging markets

Technical barriers addressed:‒ Specific improvements in metabolic pathway engineering to yield highly

productive, high yield fermentation technology‒ Novel product recovery and process integration‒ Design of new catalysts and processes (for derivatives of 3-HP)

Project Goals and Objectives

Cargill, through collaboration with their partners Codexis and PNNL will develop a new bio-based platform technology to produce a portfolio of products based on 3-HP produced by the fermentation of carbohydrates. This project will deliver an organism and process for 3-HP production, and catalysts and complete process concepts suitable for piloting and scale-up for industrial production.

Our current timeline fits well with the FY08 Solicitation schedule

O

O O

CH2OH

O

CH2NH2

O

3-hydroxypropionic acid

1,3-propanediolEEP

acrylamide

acrylic acid

malonic acid

poly(hydroxypropionate)

HO OH

O

HO OH

HO OH

OO

OH

O

OH

n

Project Strategic Fit

1994 = 1001994 = 100

Project Strategic Fit

Market dynamics have emerged to make 3-HP conversion to acrylic as an attractive alternative to propylene oxidation

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Soybean Oil Crude Oil Propylene Natural Gas Glucose Syrup

Propylene Polypropylene

Acrylonitrile

Oxo Chemicals(butanol, 2-EH)

Propylene oxide

Cumene

Isopropyl alcohol

Polygas chemicals(nonene, dodecene)

Other chemical uses

Polyurethane polyolsPropylene glycolsGlycol ethers

AdiponitrileABS/SAN resinsAcrylamideNitrile rubber

PhenolAcetone

Acrylic acid

57%

• Despite a global market of 6.6 billion pounds for acrylic acid, propylene used for acrylic production is a small portion of propylene supply, with price driven principally by polypropylene

• Growth requires one new world class plant every year

10%

4%

12%

7%

8%

4%

4%

3%

Project Strategic Fit

• Sponsoring BU within Cargill is Industrial Bioproducts, a member of the Emerging Business Platform

• This project enables a lower cost (and less price volatile) route to acrylic acid from renewable sugars. Joint analysis of production economics with an industry leader indicates a potential manufacturing cost advantage vs. propylene oxidation of >5¢ per pound for a Midwest plant.

• Energy consumption would be reduced 61% compared to the petrochemical route. By 2020, the acrylic acid product alone would displace 7,200 barrels of oil/day with domestically produced corn.

• The biorefinery route to acrylic acid would eliminate 600,000 lbs of SOx, 1 million lbs of NOx and 2.8 billion lbs of CO2 annually.

Project Strategic Fit

Project Approach

L--alaninePyruvate

Glucose

-alanine

3-HP

malonate semialdehyde

1 2 3

4

1 – Pyruvate / alanine aminotransferase2 – Alanine 2,3-aminomutase3 – -alanine aminotransferase4 – 3-HP dehydrogenase

α-ketoglutarate glutamate glutamate α-ketoglutarate

NADH

NADH

ATPH2N

O

OH

NH2

O

OH

O

OH

O

O

OH

HO

Project Collaboration

Glucose 3HPAcrylic AcidAcrylamide

Microbial catalysis/fermentation

Chemical catalysis

PNNL: Catalyst & process development

Codexis: Enzyme & pathway evolution

Cargill: Host strain construction & fermentation process

Cargill: Process design, integration & economics

Task Group 1 Task Group 2 Task Group 3

Project Tasks

Task Groups

1. Biochemical pathway optimization

2. Host strain construction and fermentation process development

3. Catalyst screening and development, process development and process integration for the conversion of 3-HP to acrylic acid

Project Tasks

Task Group 1: Biochemical pathway optimization (Codexis)

Targets• Improve performance of Enzyme 2 to meet

productivity targets• Balance supporting pathway enzymes to

maximize productivity and yield

Milestones• Performance measured by fermentation

productivity (see Task Group 2)

Technical Risks• Enzyme 2 does not meet activity target

Future plans • Continue optimization of Enzyme 2 • Perform pathway shuffling to optimize pathway performance

• Random• Phylogenetic – semisynthetic• Targeted• Hit shuffling• ProSAR

WTPar

entRd1 Rd2 Rd3 Rd4 Rd5 Rd6 Rd7 Rd8

Rounds of shuffling

Rel

ativ

e im

pro

vem

ent

Project Tasks

Task Group 2: Host strain construction and fermentation process development (Cargill)

Targets Host strain selected and pathway engineered into strain Pathway constructed and 3-HP produced• Fermentation optimization to meet economic targets

Milestones Specific productivity targets communicated to DOE

(confidential)

Technical Risks• Targets for high productivity and titer are not met

Future plans• Incorporate improved variants of Enzyme 2 into pathway• Improve yield and productivity of fermentation process

Project Tasks

Task Group 3: Catalyst screening and development, process development and process integration for the conversion of 3-HP to acrylic acid (PNNL, Cargill)

Targets Catalysts screened and selected for the dehydration of 3-HP to acrylic

acid Catalyst & process conditions demonstrated near quantitative

conversion and selectivity

Milestones • Milestones met for pure 3-HP

Technical Risks• Catalyst robustness/ effect of impurities

Future plans• Demonstrate extended catalyst life• Demonstrate catalyst performance on 3-HP produced by fermentation• Process integration

• This project enables the first biobased route to acrylic acid, a commodity product with a global market approaching 7 billion pounds per year.

Market & Customers

Water Treatment

3%

Acrylate Esters56%

SAPs32%

Miscellaneous3%

Detergents6%

• Meeting economic targets for acrylic will also enable other 3-HP derivatives.– 1,3-propanediol– malonic acid and others

• Cargill’s route to acrylic acid through 3-HP is advantaged over other technology options

– Propane oxidation is unproven and does not meet DOE OBP objectives

– Lactic acid dehydration pursued by many parties without success

– Future may allow use of biomass feedstock for Cargill process, thus driving costs downward

• Relative attractiveness of 3-HP to acrylic technology will depend on propylene price, but current forecast for high prices and volatility provides the basis for optimism

Competitive Advantage

Progress and Accomplishments

Production of 3-HP from glucose demonstrated by a novel pathway

Catalyst discovered for 3-HP to acrylic acid process

Dehydration reactor designed to overcome equilibrium limitations

Future Work

Development Stage Plans

• Due to a reduced rate of spend in FY04 and FY05, Cargill requests an extension of one year to complete the program. New proposed date for completion is May ‘07.

• Plans are to accelerate the spend over the next 18 months.

• Interim milestones and commercial plans communicated to DOE in July 2005 review (confidential).

• Our current timeline fits well with the FY08 Solicitation schedule