Systems using cell culture for production of isoprene

• US 9,121,039 B2
• Filed: 09/16/2013
• Issued: 09/01/2015
• Est. Priority Date: 09/15/2008
• Status: Active Grant

• Current Assignee: Goodyear Tire & Rubber Company
• Original Assignee: Goodyear Tire & Rubber Company, Danisco USA Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Chotani, Gopal K., McAuliffe, Joseph C., Peres, Caroline M., Whited, Gregory M.
• Primary Examiner(s): Kosson, Rosanne
• Application Number:
  US14/028,342
• Publication Number:
  US 20140073026A1
• Time in Patent Office: 715 Days
• US Class Current: 1/1
• CPC Class Codes: C12P 5/007 containing one or more isoprene units, i.e. terpenes carotenes C12P23/00
• CPC Class Codes: C12P 5/026 Unsaturated compounds, i.e. alkenes, alkynes or allenes
• CPC Class Codes: Y02E 50/30 Fuel from waste, e.g. synthetic alcohol or diesel

Abstract

The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells. The invention provides isoprene compositions, such as compositions with increased amount of isoprene or increased purity. Additionally, the invention provides methods of producing isoprene by culturing cells under conditions suitable for isoprene production while maintaining cell viability and/or metabolic activity.

Claims

1. A method of producing isoprene, the method comprising:

a) culturing cells comprising one or more nucleic acids encoding an isoprene synthase polypeptide for production of isoprene;

b) producing isoprene, wherein the cells produce greater than 400 nmole/g_wcm/hour of isoprene, and wherein the carbon dioxide evolution rate of the cells is greater than 1×

10<sup>−

18</sup> mmol/L/hour; and

c) recovering the isoprene made in step (b).

2. The method of claim 1, wherein the cells further comprise one or more heterologous nucleic acids or one or more additional copies of an endogenous nucleic acid encoding an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide, a mevalonic acid (MVA) pathway enzyme, or a deoxyxylulose-5-phosphate (DXP) pathway enzyme.

3. The method of claim 2 wherein the MVA pathway enzyme is mevalonate kinase.

4. The method of claim 1, wherein the isoprene synthase polypeptide is from Pueraria, Populus or the hybrid Populus alba×

Populus tremula.

5. The method of claim 4, wherein the source of the isoprene synthase polypeptide is selected from the group consisting of Pueraria montana, Pueraria lobata, Populus tremuloides, Populus alba, Populus nigra, and Populus trichocarpa.

6. The method of claim 1, wherein the cells are gram-positive bacterial cells, Streptomyces cells, gram-negative bacterial cells, Escherichia cells, Pantoea cells, fungal cells, filamentous fungal cells, Trichoderma cells, Corynebacterium cells, Aspergillus cells, or yeast cells.

7. The method of claim 6, wherein the cells are selected from the group consisting of Bacillus subtilis, Streptomyces lividans, Streptomyces coelicolor, Streptomyces griseus, Escherichia coli, Pantoea citrea, Trichoderma reesei, Corynebacterium glutamicum, Aspergillus oryzae, Aspergillus niger, Saccharomyces cerevisiae and Yarrowia lipolytica.