Methods of producing isoprene and a co-product

• US 8,895,288 B2
• Filed: 12/30/2009
• Issued: 11/25/2014
• Est. Priority Date: 12/30/2008
• Status: Expired due to Fees

• Current Assignee: Goodyear Tire & Rubber Company
• Original Assignee: Goodyear Tire & Rubber Company, Danisco US, Inc. (International Flavors & Fragrances Incorporated)
• Inventors: Chotani, Gopal K., Peres, Caroline M., Wells, Derek H., Sanford, Karl J., McAuliffe, Joseph C.
• Primary Examiner(s): PROUTY, REBECCA E
• Application Number:
  US12/650,332
• Publication Number:
  US 20100196977A1
• Time in Patent Office: 1,791 Days
• Field of Search: None
• US Class Current: 435/252.3
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 5/007 containing one or more isoprene units, i.e. terpenes carotenes C12P23/00
• CPC Class Codes: C12P 7/06 Ethanol, i.e. non-beverage
• CPC Class Codes: C12P 7/065 with microorganisms other than yeasts
• CPC Class Codes: C12P 7/18 polyhydric
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The invention features methods producing isoprene and a co-product, such as ethanol, 1,3-propanediol, or hydrogen from cultured cells. The invention also provides compositions that include these cultured cells. The invention provides compositions comprising isoprene and ethanol, isoprene and 1,3-propanediol, and isoprene and hydrogen. Additionally, the invention provides methods of co-producing isoprene and ethanol, isoprene and 1,3-propanediol, and isoprene and hydrogen by culturing cells under conditions suitable for co-production of isoprene and ethanol, isoprene and 1,3-propanediol, and isoprene and hydrogen.

Claims

1. Recombinant host cells comprising (a) a heterologous nucleic acid encoding an isoprene synthase polypeptide, (b) a nucleic acid encoding an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide, (c) nucleic acid(s) encoding a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide and/or one or more mevalonate (MVA) pathway polypeptides and (d) nucleic acid(s) encoding one or more propanediol pathway polypeptides,

wherein said cells co-produce isoprene and 1,3-propanediol under oxygen-limited culture conditions, wherein the oxygen transfer rate (OTR) is less than the oxygen uptake rate (OUR).

2. The cells of claim 1, wherein the heterologous nucleic acid encoding an isoprene synthase polypeptide is operably linked to a promoter.

3. The cells of claim 2, wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.

4. The cells of claim 3, wherein the plant isoprene synthase polypeptide is from Populus alba.

5. The cells of claim 1, wherein the nucleic acid(s) of (d) encode a polypeptide involved in the glycerol pathway or the 1,3-propanediol pathway.

6. The cells of claim 5, wherein the polypeptide involved in the glycerol pathway or the 1,3-propanediol pathway is dihydroxyacetone phosphate reductase (DAR1), glycerol-phosphate phosphatase (GPP2), glycerol dehydratase B1 (dhaB1), glycerol dehydratase B2 (dhaB2), glycerol dehydratase B3 (dhaB3), dhaX, orfX, orfY, 1,3-propanediol oxidoreductase (dhaT), glycerol dehydrogenase (dhaD), or dihydroxyacetone kinase (dhaK).

7. The cells of claim 1, wherein the cells further comprise one or more heterologous nucleic acid encoding a deoxyxylulose-5-phosphate (DXP) pathway polypeptide.

8. The cells of claim 1, wherein the cells comprise a DXP pathway polypeptide and a MVA pathway polypeptide.

9. The cells of claim 1, wherein the cells comprise one or more MVA pathway polypeptides selected from group:

(i) an acetoacetyl-Coenzyme A synthase (thiolase) polypeptide;

(ii) a 3-hydroxy-3-methylglutaryl-Coenzyme A synthase polypeptide (iii) a 3-hydroxy-3-methylglutaryl-Coenzyme A reductase polypeptide, (iv) mevalonate kinase (MVK);

(v) phosphomevalonate kinase (PMK);

or (vi) diphosphomevalonate decarboxylase (MVD).

10. The cells of claim 1, wherein the cells are E. coli cells.