Development of a Bio-Based

Process for Isoprene

October, 2012

David Benko, Frank Feher, and Tang Wong

The Goodyear Tire & Rubber Co.

Akron, OH

Greg Bohlmann, Greg Whited, Maggie Cervin,

Joe McAuliff, Rich LaDuca, and Karl Sanford

Genencor Division of Danisco USA

Palo Alto, CA

2

Typical Passenger Tire Composition

Synthetic &

Natural Rubber

43%

Carbon Black

& Silica

27%

Steel

10%

Fabric

5% Chemicals

10%

Oil

5%

Oil and

Natural Rubber

Oil + Energy

Oil

+

Energy

Energy

+

Raw

Materials

Over 70% petroleum derived

3

Natural Rubber (NR) is a Key Resource for Tires

• Large volumes available

– Global NR production is ~20 B lbs/yr

– Approximately 70% is used to make tires

• Superior physical properties

– Excellent resistance to wear and tear; low hysteresis

– Essential for demanding applications (e.g., aircraft,

military, trucks)

– Major ingredient (25-50%) in better consumer tires

• No alternatives available at comparable price and volume

– Commercial NR based on single species (i.e., Hevea brasiliensis)

– Synthetic polyisoprene is best alternative but limited by relative scarcity of

isoprene monomer

4

Threats to Supply of Natural Rubber

• Global demand increases well beyond supply

• Political instability in growing regions (Southeast Asia, W Africa) – Rubber trees grow in a very narrow band at +/- 12 degrees from the

equator.

• Economic instability – Dominance of small holders

– Economic alternatives from industrialization

• Widespread plant disease from lack of genetic diversity

Significant threats to NR supply exist

5

World Map Showing NR Production Regions

+ 12°

Hevea Brasilienis (The Rubber Tree) is widely distributed in the Equatorial Belt

The solid areas show the main centers of production

Rubber is a Tropical Tree (best at Equatorial zone) that grows best at temp of 24-30°C (75-86°F)

Needs annual rainfall of 1,780-2,030mm (70-80in)

Up to 610m (2,000ft) above sea level. Prime area ±12 degrees latitude

L. AMERICA

2.0%

Tropic of Cancer 22½ N

Tropic of Capricorn 22½ S

Equator

AFRICA

4.3%

- 12°

Thailand, Indonesia

& Malaysia account

for 71.3%

China

India ASIA

93.7%

Hainan, China (outside zone)

not best,

but strategic

6

The Vision: Isoprene from

Biomass

We aim to develop an efficient and sustainable fermentation

route to isoprene from carbohydrate feedstocks

2-Methyl-1,3-butadiene

Corn Switchgrass

Sugar cane

MolassesWoody

biomass

Isoprene

BiomassSugars

D-Glucose

+

other C5 and C6

sugars

O

OH

HOHO

OH OH

7

Biochemical Basis for Feedstock Conversion

CO2 + H2O

α-D-Glucose β-D-Glucose

enzymes

enzymes

enzymes

cellulose

starch

isoprene

enzymes

8

Bioprocess to Isoprene Enables:

• Decreased dependence on natural rubber sourcing

• Reduced petroleum price impact on isoprene costs

• Supply of isoprene de-linked from petroleum processing

• Stability of monomer supply

• Potential for significant savings associated with rubber supply

• Potential for product performance enhancement

Synthetic rubber from sustainable nonpetroleum based sources

9

Isoprene Production

TREES BIOBASED

PROCESS

CHEMICAL

SYNTHESIS

PETROLEUM

CRACKING

• Slow

• Not commercially relevant

• Extractive distillation from ethylene & propylene production

• Price linked to oil prices

• Moving to “light” streams

• High cost

• Petroleum feedstocks

• Mfg. infrastructure in Russia & Japan

• Renewable feedstocks

• Biochemical and biofuel products

10

Building Block Biochemicals (2006):

11

Partnership for BioIsopreneTM

• Leading manufacturer of synthetic elastomers

• Technology for producing synthetic polyisoprene

• Expertise for isolating, purifying, handling, storing and shipping isoprene

• Leading industrial biotechnology

company

• Technology for producing

genetically-modified organisms for

industrial applications

• Ability to develop complete process

for producing isoprene from

biomass

Collaborative research initiative with joint multimillion-dollar investments

12

Goodyear and Genencor leveraging each

others’ expertise …

CH2

CC

H2C

CH3H

Integrated biobased process for production of BioIsopreneTM

Renewable Feedstock(s)

Microbial Strain Development

Large Scale Fermentation

BioIsopreneTM recovery & purification knowhow

Polymer-Grade BioIsopreneTM

Product

13

BioIsopreneTM Technology:

• Multiple feedstocks possible

• Two feeder pathways to isoprene precursor

• Isoprene synthase

C6 3C (C5)

3C 2C

16C 2C

Isoprene Synthase

Isoprene Recovery

Glucose

Sucrose / Biomass

Glycerol

Plant Oils

DXP Pathway

MVA Pathway

Cells

Conceptual View of Cell Factory

14

Metabolic Pathways

Bouvier (2005) Prog. Lipid Res. 44, 357–429.

• Two known pathways for

producing precursors to

isoprene (MVA, DXP)

• Both capable of using

fermentable sugars as

feedstocks

• Same isoprene-producing

reaction

15

Genetic Engineering Optimizes Bacteria for Isoprene Production

Enterococcus

Yeast

Archea

Kudzu

E. Coli

Genes from 5 different organisms were used to create the 1st-

generation isoprene-producing bacteria

16

• Engineered E. coli

• 14-L fed batch fermentation from glucose

• Extensive IP portfolio: 14 published applications to date

BioIsoprene™ Titer

0

50

100

150

200

250

0 8 16 24 30 37

Time (h)

OD

600

0

10

20

30

40

50

60

70

Tit

er

(g/L

)

Jan 2009

Titer

OD600

Isoprene Production

17

Integrated Process Overview

PF

FF

PF

FF

L

FR

P

I

P

R

P

R

Polymerization Fermentation Recovery - Purification Feedstock

BioIsoprene™

Product

BioNatsyn™ (Synthetic

Rubber)

18

BioIsopreneTM Polymerization

Frequency Sweep of Tan d for PetroNatsyn & BioNatsyn

0.0

0.1

0.2

0.3

0.4

0.5

1 10 100 1000 10000

Frequency (cpm)

Tan

d

Petro Natsyn

BioNatsyn

19

Tread Compound Properties

BioNatsyn

Polyisoprene Natsyn Polyisoprene

Cure time, min

(t90 at 150°C)

19.6 21.8

300% modulus,

MPa

9.2 7.2

Tensile strength,

MPa

16.5 16.1

Elongation at

break, %

542 598

Mooney viscosity 45 45

Shore A hardness 71 62

The recipe consisted of 100 phr cis-1,4-polyisoprene, 60 phr carbon black, 12.5 phr

silica, 13 phr oils and waxes, 8.25 phr antidegradants, 10.8 phr curatives

20

Concept Tire

21

2010 - Add isoprene to the list