Genomatica Sustainable Chemicals

Genomatica Sustainable Chemicals

Development of an Integrated Biofuel and Chemical Refinery

John D. Trawick

Research Fellow, Genomatica

2013 DOE Bioenergy Technologies Office (BETO) Project Peer Review

Date: 21 May 2013

Technology Area Review: Biochemical Conversion

Principal Investigator: Mark Burk Organization: Genomatica

This presentation does not contain any proprietary, confidential, or otherwise restricted information

Goal Statement Demonstrate the viability and commercial readiness of an integrated

biorefinery for low cost production of 1,4-butanediol (BDO), from biomass—deliver the engineered strain and optimized fermentation

process to enable the conversion of cellulosic sugars into BDO.

1: Improving the microbial conversion of cellulosic sugars to BDO.

To deliver commercially acceptable performance and enable scalable integrated biorefineries.

2: Characterizing and improving tolerance to cellulosic hydrolysate.

To deliver commercially acceptable performance and enable scalable integrated biorefineries.

3: Developing and optimizing a scalable fermentation process. Demonstrate the feasibility and scalability of integrated biorefineries.

To deliver a scalable fermentation process that employs the engineered microbe to produce BDO from cellulosic sugars at titer ≥ 70 g/L, and productivity ≥ 2.0 g/L/hr at ≥ 100 L scale.

Quad Chart Overview

• August 2011 • July 2014 • ~80% complete

• Barriers addressed • Consistency, quality, and concentration of

cellulosic sugars in hydrolysates. • Glucose – Xylose – Arabinose co-utilization • BDO T-R-Y metrics in hydrolysates vs.

refined sugar

Funding for FY11(541,971/186,308) Funding for FY12(852,555/293,074) Funding for FY13 (253,588/87,173) 2/ $1,000,000.

Timeline

Budget

Barriers

• Chemtex • Suppliers of PROESATM hydrolyates, have worked

with Genomatica to reach a specification • Biweekly or more frequent consultation with

Chemtex staff

Partners

Project Overview Genomatica has developed recombinant organisms to produce the commodity, 1,4-butanediol (BDO), used in many synthetic polymers.

•BDO producing strains use refined dextrose to make BDO; ties BDO economics to sugar and corn prices.

•Biomass-based sugars are an economical alternative.

•Challenges include:

o Cheap, consistent feedstock and treatment.

o Minimizing hydrolysate impurities.

o Achieving organism performance metrics: titer (T), rate (R), and yield (Y) similar to dextrose-based.

o Simultaneous utilization of C5 as well as C6 sugars by recombinant Escherichia coli.

o Recovery of BDO is different than ethanol and this impacts design and economics.

1 - Approach • E. coli to make 1,4-BDO at titer, rate, and yield from lignocellulosic

biomass sugars to demonstrate commercial feasibility.

• Critical factors: 1) hydrolysate quality/composition, 2) C5-C6 co-utilization, 3) process impacts on yield and rate.

• Hydrolysate composition, work with supplier to produce biomass hydrolysates with high and uniform [sugar], low impurities, and minimal toxicity.

– Go/no go: achieve 75% T-R-Y of pure glucose at same concentration.

• Adaptive evolution + genomic re-sequencing for sugar co-utilization.

– Go/no go: Efficient C5-C6 co-utilization and move to ‘clean’ strain

• 13C flux analysis + metabolomics to ID metabolic constraints limiting performance.

– Targets to improve energy and reduced cofactors.

Bio-based 1,4-Butanediol

Crude Oil & Nat. Gas

Acetylene

Butane, Butadiene

Propylene

2.8 B lb/yr existing market

spandex polyurethanes

PBT

Sugars

Direct Production

Genomatica’s Systems-Based Strain Engineering

Whole Cell Mutagenesis

Metagenomics, Enzyme Evolution

Computational Technologies

Pathway &

Strain Design

Product Feedstock Organism & Tools Select

Parent strain

Engineered strains

HT Screening In vivo assays

Metabolic Engineering Tools HT Cloning

Dat

a

LIMS Fermentation development/ scale-up

Omics data Systems analysis

Analyze & Interpret

13C-Fluxomics

Metabolomics

Proteomics

Transcriptomics

Genomics

Iterative Strain

Engineering

Biological

Knowledge

Journey to a BDO Production Strain

Pathway Identification and Engineering

glucoseex

acetyl-CoA citrate

phosphoenolpyruvate

glyoxylatemalate

pyruvate

ADP, NAD+

ATP, NADH

NAD(P)HCO2

NADH CO2

NAD+

ATPCoA

ADPPi

succinate

oxaloacetate

succinyl-CoA

fumarate

NAD+

NADH,CO2

CO2

hexose-P

-ketoglutarate

NAD(P)+

CoA

succinatesemialdehyde

CO2 NAD(P)+NAD(P)H

isocitrate

ATP

ADP

4-hyd

roxy

butyrate

4-hyd

roxy

butyrylC

oA

acetate

AMP

quinol

quinone

4-hydroxybutyryl-aldehyde

1,4-butanediol

1,4-butanediolex

NAD(P)+

NAD(P)H

NAD(P)+

, CoA

ATP, CoA

NAD(P)H

sucC

D

sucA

sucD

4hbd

ald

adh

PTS system

ppc

ATPADP

sucroseex

glk

ATP

ADP

frk

NAD(P)+

NAD(P)H

acs

sucAB, lpdA

lpdA

acetyl-CoA

mqoicdA

fumABC

sdhABCD

acn

aceAaceB

gltA

ubiquinone

ubiquinol

pckA

ADPCO2

ATP

cat2

renewable feedstock

sustainable chemicals

mdh

arcA

ldhadhEpflB

K.p.lpdD354KlpdA

gltAR163L

rrnC::cscAKB

Strain Design and Metabolic Engineering

Commercial Strain for BDO Production

• Titer (g/L) - Impacts equipment sizing and energy needs • Rate (g/L/h) - Impacts # of fermentors, plant capacity • Yield (g/g) - Impacts feedstock cost contribution

Fermentation Metrics → Higher TRY = Lower COGS

TRY all inter-dependent → reduce by-products, increase rate and yield

BDO Biosynthetic Pathways

Alternative routes

1. Developed enzyme assays and analytical methods for all metabolites 2. Screened libraries of gene candidates for each step – >100 in some cases 3. Demonstrated seven different functional BDO pathways in E. coli

Sugar

• Prioritized pathways proceed through 4-hydroxybutyrate • Downstream enzymes function on non-natural substrates

ALD Cat2 ADH

Yim et al., Nature Chem. Biol., 2011

BDO Pathway and Process

• BDO pathway involves 4 reduction steps – redox intensive

• BDO pathway generates 1 extra NAD(P)H and no excess ATP

• Balance energy, redox and maintain high NAD(P)H/NAD(P)+ ratio

• Microaerobic production (DO ≈ 0) required for optimal performance

ATP = 0 NAD(P)H = +1

Oxidative TCA cycle flux required for redox needs of BDO pathway

ATP via oxidative phosphorylation

E. coli

Glycolysis

TCA Cycle

BDO Pathway

BDO

E. coli

Sugars Glycolysis

TCA Cycle

BDO Pathway

C6H12O6 + 0.5 O2 C4H10O2 + 2 CO2 + H2O Max yield = 1 mol/mol (0.50 g/g, 67 C-mol %)

>100 g/L

Technology Platform Drives BDO Strain Performance

3.3 g/L-h 2.5 g/L-h 2.0 g/L-h

2011

2012

Increasing Rate and Lowering By-products

More metabolic steps in a pathway increases avenues for by-products

Key Advances • Backflux • Enzymes • Redox • ATP supply • Regulation • Balanced expression • Fermentation PD

2 PEP

PYR

Ace ACCOA OA

G6P

CIT

ICIT

AKG

SUCSAL

4HB

4HBALD

BDO

+ATP

-ATP

+NADH

+NADPH

- NADH

- NAD(P)H

+2 NADH

- NAD(P)H

1 GLUCOSE

CO2 loss

Acetate

Glutamate

CO2 loss via oxidative TCA

4-HB

Ethanol -2 NADH

SUCCOA

TCA Cycle

GBL

+ATP

Metabolic Engineering Applications

Approach to Biomass - BDO • Initial baseline performance (preliminary results in the grant).

• Identification of key constraints on Biomass – BDO and overcoming those constraints

o Hydrolysate composition, work with supplier to produce biomass hydrolysates with high and uniform [sugar], low [impurities], and minimal toxicity. o Hydrolysate improvements with time increased performance.

o Adaptive evolution + genomic re-sequencing + strain engineering for sugar co-utilization. o Acceptable sugar co-utilization selected, designed, and recapitulated.

o 13C flux analysis + metabolomics to ID metabolic constraints limiting performance. o Targets to improve availability of energy and reduced cofactors.

Benchmark performance on biomass, September 2011

• Benchmark fermentations for DOE/NREL site visit using early lot of hydrolysate and a very early BDO production strain*.

• Titer (16 – 18 g BDO/L) was 24% of grant goal and rate (0.25 g/L/hr) <10% of goal.

• Yield (not shown) >50% of goal.

• A long way to go!

*As used in grant application.

Biomass-to-BDO Process Challenges: impurity reduction

35% Glucose 007, Chemtex Hz, low impurities 001, Chemtex Hz, diluted w/sugar 001, Chemtex Hz, untreated Benchmark, 9/’11, early strain, Hz similar to 001 Strain: BDO Producer Chemtex Hydrolysate at 35% sugars in feed: • Lot 001, Untreated Hz • 001, Untreated Hz

Diluted with pure glucose + xylose

• Lot 007: Hz

Chemtex provided hydrolysates to evaluate •Sugar concentration in a fed-batch process—limits titer and rate •Lowered impurities; reduced multiple ways

Result: BDO titer approached metric on pure glucose Strain improvements from original benchmark coupled with Hz improvements.

BDO titer (g/L)

Biomass-to-BDO Process Challenges : Impurity reduction

35% Glucose Chemtex Hz, low impurities Chemtex Hz, diluted w/sugar Chemtex Hz, untreated Benchmark, 9/’11, early strain, Hz similar to 001 Strain: BDO Producer ChemtexHydrolysate at 35% sugars in feed: •Untreated •Diluted with pure glucose + xylose •Low impurities

Chemtex provided hydrolysates to evaluate •Sugar concentration in a fed-batch process—limits titer and rate •Lowered impurities; reduced multiple ways

Result: BDO titer approached metric on pure glucose Strain improvements from original benchmark coupled with Hz improvements.

BDO rate (g/L/hr)

Escherichia coli BDO production strains show diauxic sugar utilization

Compared, wt MG1655 vs. a BDO production strain 50/50 glucose xylose, growth curves

Conclusion: BDO production strains have lessened diauxie

relative to the wt progenitor.

0.000

0.100

0.200

0.300

0.400

0.500

0.600

0.700

0

2.2

5

4.5

0

6.7

5

9.0

0

11

.25

13

.50

15

.75

18

.00

20

.25

22

.50

24

.75

27

.00

29

.25

31

.50

33

.75

36

.00

38

.25

40

.50

42

.75

45

.00

0.1% glucose + 0.1% xylose2731/pZE13

0.1% glucose + 0.1% xylose MG1655

Using Evolution to Improve Xylose Sugar Utilization in Fermentation

Parent unevolved parent BDO producing strain.

Evolved : evolved from Parent for improved xylose utilization. Fermentation used pure glucose : xylose (1:1), 680 g sugar/L feed

Selection gave a large improvement in xylose consumption in the presence of glucose

• Fermentation in 2 L BR • Strains made BDO in glucose + xylose;

• Evolved = 70 - 75 g/L; • Parent = 60 g/L

0%

20%

40%

60%

80%

100%

0 10 20 30 40 50

% G

luc/

Xyl

C

on

sum

ed

Time (hrs)

%Glucose/Xylose Consumption

Evolved Glucose Cons

Evolved Xylose Cons

Parent Glucose Cons

Parent Xylose Cons

0

40

80

120

160

200

240

0 10 20 30 40 50

Tota

l Glu

c/X

yl C

on

sum

ed

(g)

Time (hrs)

Total Glucose/Xylose Consumption

Evolved Glucose Cons

Evolved Xylose Cons

Parent Glucose Cons

Parent Xylose Cons

Evolved and re-capitulated for glucose + xylose co-utilization

• XUM (xylose utilizing mutant) for xylose co-utilization with glucose was

identified and integrated into a clean BDO production host. • A gene for arabinose uptake also added. Result: Efficient co-utilization of all 3 sugars during fermentation.

0.00

20.00

40.00

60.00

80.00

0 8 13 18 23 28 33

Re

sid

ual

Su

gar

(mM

)

Elapsed time (hours)

C6 - C5 Co-Consumption

Glucose

Xylose

Arabinose

Residual [arabinose] in non-arabinose utilizing

strains with similar biomass hydrolysates for

comparison.

Improving BDO Performance on lignocellulosic hydrolysates with strain engineering and hydrolysate characteristics

ID C u ltu re ID Eq u ip Start Date In i t OD Tite r P ro d Yie ld C arb o n B alan ce

IR R Time p o in t Yie ld B asis C ate go ry Varyin g C o mp o n en t

LIM S ID Strain Name Strain Al t Name

No me n clatureP lasmid R e vie we d Ou tl ie r

1 4 8 7 -1 B R 1 7 4 /1 0 /2 0 12 1 2 :0 0 :00 AM

3 .9 2 3 9 .2 0 3 0 .8 9 1 0 .2 4 2 8 5 .8 3 7 6 .8 4 4 [Ara], [Glu c], [X yl ]

P ro ce ss De ve lo p men t

1 0 % M &G WS Hyd ro lysate , M M 9 , 2 mM

M gSO4, 2x TE , M &G WS Hyd ro lysate C e n tri fu ged an d Fi l te re d

(0 0 1-12-WS-4X)

L4 3 2 1 EC K h -3 9 33 + p ZS* 1 3S-dBsaI-7 1 4 B K -143 8A

Ed n a p ZS* 1 3S-dBsaI-7 1 4 B K -143 8A

N N

2 6 8 9 -1 B R 2 9 /2 5 /2 0 12 1 2 :0 0 :00 AM

5 .6 8 7 6 .7 9 1 .6 0 .4 8 6 1 5 5 .8 82 1 7 .5 4 8 [Ara], [Glu c], [X yl ]

Strain Scre e n in g

FM 8 - 3 .65 % M &G Lot 012 , M &G-0 1 2-WS-

NH3 , Raptor 1 o n Lo t 0 12

L5 8 4 1 R ap to r 1 re tran sfo rmed

R ap to r 1 re tran sfo rmed

N N

3 7 6 9 -1 B R 1 1 1 2 /4 /2 0 12 1 2 :0 0 :00 AM

3 .6 8 9 .1 1 5 1 .7 3 0 .3 9 3 1 2 0 .2 33 1 6 .3 2 5 1 .5 [Ara], [Glu c], [X yl ]

Strain Scre e n in g , M e d ia

De ve lo p men t

FM 2 5 - 2 .88% M &G Lot 010 , M &G-0 1 0 ,12-

WS-S-NH3 , Lot 0 1 0

L6 2 0 2 EC K p -5 1 57 + p ZS* 1 3 S-p246-1 4 3 8 -14 39-

1 6 0 B-p 115 -3 0 1 -3 02

EC K s-5 4 1 1 p ZS* 2 3 S-p246-1 4 3 8 -14 39-1 6 0 B -p 115 -

3 0 1 -3 02

N N

• Better strains and biomass treatment = improved BDO titer and rate

• Early 2012 diauxic strain, early Hz, Chemtex

• Late ‘12, Evolved/recapitulated xylose user, Chemtex, low impurities

• Late ‘12, Evolved/recapitulated xylose user, Chemtex, low impurities

• >2X in increase in titer, and in peak rate during 2012

• Challenges remain • Consistency/sugar concentration • Reducing non-fermentables

Non-evolved strain Chemtex Hz-001, early batch Recapitulated, XUM Chemtex Hz-012, improved Recapitulated, XUM Chemtex Hz-010, improved

BDO titer (g/L) Final titer, 89 g

BDO/L

Final titer, 85 g BDO/L

Final titer, 39 g BDO/L

BDO rate (g/L/hr)

Cellulosic biomass (xylose) and metabolism

ADP

sucroseex

ATP

frk

acetyl-CoA

citrate

phosphoenolpyruvate

glyoxylatemalate

pyruvate

NADPHCO2

NADH CO2

NAD+

ATPCoA

ADPPi

succinate succinyl-CoA

fumarate

NAD+

NADHCO2

CO2

-ketoglutarate

NAD+

CoA

succinatesemialdehyde

glutamate4-aminobutyrate NAD+NADH

isocitrate

4-hydroxybutyrate

4-hydroxybutyrylCoA

g-butyrolactone

acetateCO2

ADP

CoAquinol

quinone

4-hydroxybutyrylaldehyde

1,4-butanediol

1,4-butanediolex

NAD(P)+

NAD(P)H

NAD(P)+

, CoA

ATP, CoA

NAD(P)H

sucC

D sucD (035)

4hbd (036)

cat2 (033U)

ald ( 714B)

adh (956)

ppc

mdh

acetyl-P

NADP+

NADH

ack,pta

sucAB, lpdA

lpdA

acetyl-CoA

1,4-Butanediol Biosynthetic Pathway

mqo

NAD(P)HCO2

NADP+

sfcA,maeB

icdA

fumABC

sdhABCD

R163Lacn

aceAaceB

gltA

ubiquinone

ubiquinol

pepck

ADPCO2

ATP

Confidential

aspAaspartate

aspC

glutamate-ketoglutarate

car (891)

NADP+

NADPH

±

±

+ pptase (910)

glutamateNADP+

NADPH

gdh

glutamateex

gltBD

glucoseex

ADP, NAD+

ATP, NADH

fructose6-P

ADP

PTS system

ATPADP

glk

fructose 1,6-P

fbp pfkABADP

ATPPi

pgi

glucose 6-P

glyceraldehyde 3-PfbaAB

PEP

pyruvate

pykAF

zwf

ATP

esterase

4-hydroxybutyrateex

ribulose 5-P

xylulose 5-P

ribose 5-P

sedoheptulose 7-P

erythrose 4-P

NADPHNADP+

ripAB rpe

tktAB

tktAB

talB

NADPH,CO2NADP+

gnd

pgl

6P-gluconolactone

6P-gluconate

gabT

gadAB

xylB

xylA

xyloseex

xylulose

xylosexylEFGH

sucroseex

glucose

fructose

permease

invertase

ADP

ATP

CoAPi

pta

acetate

ATP

ADP

acetateex

ackA

oxaloacetate

13C flux analysis, metabolomics, and metabolic modeling used to ID energy

and metabolic constraints on BDO production in hydrolysates with mixed

sugars.

Including,

Net ATP reduced on xylose because of increased transport cost from xylFGH ABC

transporter

Key Accomplishments and challenges • Improved hydrolysate composition—addresses impurities, sugar

concentrations

• Glucose – Xylose – Arabinose co-utilization

• These and other changes have given at small scale fermentation:

o Chemtex hydrolysates: 86 – 89 g BDO/L titer, 1.8 g/L/hr rate

o DOE grant goal (30L scale): 70 g BDO/L titer, 1.5 g/L/hr rate

• Potential drains on energy and reducing power identified via metabolomics, 13C flux analysis, and metabolic modeling.

• Have multiple proposed changes in process of testing/implementation

3 - Relevance • Develop biomass-to-1,4-butanediol (BDO) increase chemical production

from sustainable feedstocks in a cost advantaged process that will be competitive with petrochemicals

o Achieve performance sufficient for commercial viability (BDO titer of ≥ 70 g/L, and rate ≥ 2.0 g/L/hr) Grant Tasks 1 & 2

o Scalable process to integrate into biomass based biorefineries (at ≥ 100 L scale) Grant Task 3

o Recovery of BDO along with process that is cost competitive. Grant Tasks 1, 2, & 3.

• Application of the expected outputs: Bio-BDO from cellulosic biomass, cost advantaged, that can be incorporated into the integrated biorefinery concept.

4 - Critical Success Factors • Success factors defining technical and commercial viability.

BDO titer of ≥ 70 g/L, and rate ≥ 2.0 g/L/hr at ≥ 100 L scale. o Robust organism meeting these targets (technical); >75% of the goal o From biomass, BDO purity equal to industry needs (market); can do. o Costs comparable or less than pure glucose “Gen 1 BDO” process (business);

working towards this with improved strains, hydrolysates, process.

• Potential challenges (technical and non-technical). o Cellulosic hydrolysate consistency/specs for optimum fermentation and

downstream recovery. o Strain performance in hydrolysates, especially energy cost to cell affecting yield

and productivity.

• Advance the technology and impact the commercialization of biomass/biofuels. o BDO strain/process/recovery are all distinct from ETOH or biofuels; will be a new

opportunity. o Commercially viable strain and process, biomass-to-BDO, will be very attractive to

Genomatica customers. o Begin to set specifications for biomass sugars for chemical processes

5. Future Work • Improve yield and incorporate sugar co-utilization into multiple strains. • Go/no go reaching T-R-Y targets at both 2 L and larger scale • Test new lignocellulosic hydrolysate feedstocks and treatments.

8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7

= Deliverables = Major Stage Gates

Project Month 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36

1 Improve the microbial conversion of cellulosic sugars to BDO

1.1 Maximize overall sugar utilization rate 1.1.1 Eliminate diauxic sugar utilization 1.1.2 Improve individual sugar utilization rates 1.1.3 Enable breakdown of cellobiose 1.2 Maximize BDO yield 1.2.1 Balance BDO pathway expression/flux with upstream metabolism 1.2.2 Reduce yield loss to overflow byproducts

2 Characterize and improve tolerance to cellulosic hydrolysate

2.1 Characterize and evolve tolerance to hydrolysate 2.1.1 Determine maximum tolerance to hydrolysate 2.1.2 Evolve wild-type and BDO production organism for improved tolerance 2.1.3 Characterize/resequence evolved organisms 2.2 Develop hydrolysate-tolerant production strain 2.2.1 Evaluate BDO production capabilities of evolved ‘production’ strain 2.2.2 Introduce observed mutations into unevolved production strain and

evaluate hydrolysate tolerance/BDO production

3 Develop fermentation process that consistently produces BDO from

cellulosic sugars at the target titers, yields, and productivities

3.1 Develop efficient and scalable fermentation process 3.1.1 Establish new baseline with latest BDO production strain 3.1.2 Determine maximum starting hydrolysate concentration 3.1.3 Perform initial media and protocol optimization 3.1.4 Determine effective fed-batch feeding strategy 3.1.5 Perform process development with emphasis on oxygen supply 3.2 Test improved strains and modify fermentation process as needed 3.3 Complete technology transfer package

2012 2013 2014

Task Description 2011

40 g/L, 0.75 g/L/hr, 2 - L scale

70 g/L, 1.5 g/L/hr, 30 - L scale

70 g/L, 2.0 g/L/hr, 100 - L scale

Simultaneous C5 - and C6 - sugar utilization achieved.

Growth of evolved strains in 180 g/L hydrolysate demonstrated . Strain resequencing complete.

Production strain tolerates 180 g/L hydrolysate.

Sugar utilization exceeds 4 g/L/hr. Yield goal at 2 L scale. Cellobiose utilization demonstrated.

Sugar utilization exceeds 5 g/L/hr. ? 0.42 g/g achieved at 2 - L scale.

Summary 1) Approach

1) Strain design and engineering coupled with adaptive evolution and ‘omics technologies to ID key constraints on feedstock quality and price.

2) Hydrolysate evaluation and improvement to reach specification.

2) Accomplishments 1) 5X improvement in titer; rate and yield are approaching goals.

2) Major C6 and C5 sugars can be co-utilized to max yield and performance

3) Key metabolic constraints ID’d

4) Working towards hz specification

3) Relevance 1) Enable commercial bio-BDO from lignocellulosic biomass sugars

4) Critical Success factors and challenges 1) Biomass sources meeting needed fermentation and BDO recovery specifications.

5) Future Work 1) Yield and pathway enhancements to reach or exceed grant metrics.

6) Technology transfer

Bio-BDO commercial progress

50 runs 120+ trucks

BASF, world’s leading BDO producer licensed to produce renewable 1,4-BDO using Genomatica’s process

Toray Industries, Inc. produced partial bio-PBTs from bio-BDO at same specs as petro-PBT

Bio-BDO Scale-up, 3 yrs to 5 M-lbs

Genomatica Biomass to BDO Contributors Molecular Biology

John Trawick

Ewa Lis

Joseph Warner

Chemtex

Alberto Ceria

Piero Ottonello

Francesco Cherchi

Kevin Gray

Microbiology

Carla Risso

Jesse Wooton

Jonathan Joaquin

Analytical Sciences

Julia Khandurina

Rosary Stephen

Lucy Zhao

Ahmed Alanjary

Blanca Ruvalcaba Rainer Wagester Korki Miller

Process Engineering

Michael Japs

Janardhan Garikipati

Fasil Tadesse

Matt White

Christophe Schilling, CEO Mark Burk, CTO Bill Baum, CBO Nelson Barton, VP R&D Jeff Lievense, EVP, Process Development

Computational

Tony Burgard

Priti Pharkya

Robin Osterhout

Jun Sun

Tae Hoon Yang

Jungik Choi

Harish Nagarajan

Fermentation

Dan Beacom

Sy Teisan

Laurie Romag

Joseph Woodcock

Gian Oddone

Amruta Bedekar

Jason Crater

Akhila Raya Award DE-EE0005002 to Genomatica

Questions?

Thank you John Trawick jtrawick@genomatica.com

Publications, Presentations, and Commercialization

• Barton, Nelson (VP, R&D, Genomatica) Biomass 2012, 11 – 12 July 2012, Wash., DC http://www1.eere.energy.gov/biomass/pdfs/bio2012_final_agenda.pdf

• Trawick, John D. (Research Fellow, Genomatica) scheduled for the 2013 SBFC meeting (2 May 2013) http://sim.confex.com/sim/35th/webprogram/Session2437.html

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Responses to Previous Reviewers’ Comments

• N/A.

Note: This slide is for the use of the Peer Review evaluation only – it is not to be presented as part of your oral presentation, but can be referenced during the Q&A session if appropriate. These additional slides will be included in the copy of your presentation that will be made available to the Reviewers and to the public.