# US 2014/0155660 A1 · Compositions and methods for producing isoprene free of c5 hydrocarbons under decoupling conditions and/or safe operating ranges

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- Number: US 2014/0155660 A1
- Document ID: US20140155660A1
- Status: Published application
- Kind: A1
- Application number: 13/898,320
- Filed: 2013-05-20
- Published: 2014-06-05
- Earliest claimed: 2008-07-02
- Assignees: The Goodyear Tire & Rubber Company; Danisco US Inc.
- Applicants: The Goodyear Tire & Rubber Company; Danisco US Inc.
- Inventors: Anthony R. Calabria; Marguerite A. Cervin; Gopal K. Chotani; Richard La Duca; Joseph C. McAuliffe; Michael C. Miller; Timothy A. Sabo; Karl J. Sanford; Erin L. Spring; Gregory M. Whited
- Web page: https://patentgrep.com/patent/US20140155660A1

## CPC

- C12P 5/007: {containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00)}
- C07C 11/18: Isoprene
- C12N 9/88: Lyases (4.)
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12N 15/70: Vectors or expression systems specially adapted for E. coli
- C12N 15/74: Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N 15/75: for Bacillus
- C12N 15/80: for fungi
- C12P 5/02: acyclic {(C12P5/007 takes precedence)}
- C12Y 402/03027: Isoprene synthase (4.2.3.27)
- Y02E 50/30: Fuel from waste, e.g. synthetic alcohol or diesel
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C07C: ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

The invention features methods for producing isoprene from cultured cells wherein the cells in the stationary phase. The invention also provides compositions that include these cultured cells and/or increased amount of isoprene. The invention also provides for systems that include a non-flammable concentration of isoprene in the gas phase. Additionally, the invention provides isoprene compositions, such as compositions with increased amount of isoprene or increased purity.

## Claims

### Claim 1

A biologically produced isoprene composition wherein said composition comprises greater than or about 99.90% isoprene by weight compared to the total weight of all C5 hydrocarbons in the composition.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the composition comprises greater than or about 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 mg of isoprene.

### Claim 3 (depends on 1)

The composition of claim 1, wherein the composition comprises a volatile organic fraction comprising greater than or about 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 g of isoprene (w/w).

### Claim 4 (depends on 1)

The composition of claim 1, wherein the composition comprises greater than or about 99.92, 99.94, 99.96, 99.98, or 99.99% isoprene by weight compared to the total weight of all C5 hydrocarbons in the composition.

### Claim 5 (depends on 4)

The composition of claim 4, wherein the isoprene is recovered from an off-gas portion.

### Claim 6 (depends on 1)

The composition of claim 1, wherein the composition has a relative detector response of greater than or about 99.90 for isoprene compared to the detector response for all C5 hydrocarbons in the composition.

### Claim 7 (depends on 1)

The composition of claim 1, wherein the composition further comprises less than or about 50, 40, 30, 20, 10, 5, 1, 0.5, 0.1, 0.05, 0.01, or 0.005 μg/L of a C5 hydrocarbon other than isoprene.

### Claim 8 (depends on 7)

The composition of claim 7, wherein the C5 hydrocarbon other than isoprene is selected from the group consisting of 1,3-cyclopentadiene, cis-1,3-pentadiene, trans-1,3-pentadiene, 1-pentyne, 2-pentyne, 1-pentene, 2-methyl-1-butene, 3-methyl-1-butyne, trans-piperylene, cis-piperylene, pent-4-ene-1-yne, trans-pent-3-ene-1-yne, or cis-pent-3-ene-1-yne.

### Claim 9 (depends on 1)

The composition of claim 1, wherein the composition comprises less than 5, 0.1, 0.05, 0.01, or 0.005 μg/L of a protein or fatty acid.

### Claim 10 (depends on 9)

The composition of claim 9, wherein the protein or fatty acid is naturally associated with natural rubber.

### Claim 11 (depends on 1)

The composition of claim 1, wherein the composition comprises less than or about 10, 5, 1, 0.8, 0.5, 0.1, 0.05, 0.01, or 0.005 ppm of alpha acetylenes, piperylenes, acetonitrile, or 1,3-cyclopentadiene.

### Claim 12 (depends on 1)

The composition of claim 1, wherein the composition comprises less than or about 5, 1, 0.5, 0.1, 0.05, 0.01, or 0.005 ppm of sulfur or allenes.

### Claim 13 (depends on 1)

The composition of claim 1, wherein the isoprene composition comprises less than or about 30, 20, 15, 10, 5, 1, 0.5, 0.1, 0.05, 0.01, or 0.005 ppm of all acetylenes.

### Claim 14 (depends on 13)

The composition of claim 13, wherein the acetylenes are selected from the group consisting of 1-pentyne, 2-butyne, 2-methyl-1-buten-3-yne (2 MB1-3yne), and 1-pentyne-4-yne.

### Claim 15

A biologically produced isoprene composition wherein said composition comprises less than or about 0.5 μg/L per compound for any compound in the composition that inhibits the polymerization of isoprene.

### Claim 16 (depends on 15)

The composition of claim 15, wherein the composition that inhibits the polymerization of isoprene comprises less than or about 0.12, 0.10, 0.08, 0.06, 0.04, 0.02, 0.01, 0.005, 0.001, 0.0005, 0.0001, 0.00005, or 0.00001% by weight compared to the total weight of all C5 hydrocarbons in the composition.

### Claim 17 (depends on 16)

The composition of claim 16, wherein the composition that inhibits the polymerization of isoprene comprises 1,3-cyclopentadiene, cis-1,3-pentadiene, trans-1,3-pentadiene, 1-pentyne, 2-pentyne, 1-pentene, 2-methyl-1-butene, 3-methyl-1-butyne, trans-piperylene, cis-piperylene, pent-4-ene-1-yne, trans-pent-3-ene-1-yne, or cis-pent-3-ene-1-yne.

### Claim 18

A biologically produced isoprene composition wherein said composition comprises (a) greater than about 2 mg of isoprene and (b) one or more compounds selected from the group consisting of ethanol, acetone, C5 prenyl alcohols, and isoprenoid compounds with 10 or more carbon atoms.

### Claim 19

A biologically produced isoprene composition wherein said composition comprises (a) greater than about 2 mg of isoprene and (b) one or more compounds selected from the group consisting of 2-heptanone, 6-methyl-5-hepten-2-one, 2,4,5-trimethylpyridine, 2,3,5-trimethylpyrazine, citronellal, acetaldehyde, methanethiol, methyl acetate, 1-propanol, diacetyl, 2-butanone, 2-methyl-3-buten-2-ol, ethyl acetate, 2-methyl-1-propanol, 3-methyl-1-butanal, 3-methyl-2-butanone, 1-butanol, 2-pentanone, 3-methyl-1-butanol, ethyl isobutyrate, 3-methyl-2-butenal, butyl acetate, 3-methylbutyl acetate, 3-methyl-3-buten-1-yl acetate, 3-methyl-2-buten-1-yl acetate, (E)-3,7-dimethyl-1,3,6-octatriene, (Z)-3,7-dimethyl-1,3,6-octatriene, and 2,3-cycloheptenolpyridine; wherein the amount of the one or more compounds relative to the amount of the isoprene is greater than or about 0.01% (w/w).