IP Library Granted Patent US 7,985,567
Granted Patent B2
US 7,985,567 · App. 12/646,189 · Granted Jul 26, 2011

Host cells and methods for producing 3-methyl-2-buten-1-ol, 3-methyl-3-buten-1-ol, and 3-methyl-butan-1-ol

Assignee: The Regents of the University of California
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Quick Facts
Patent No.
US 7,985,567
App. No.
12/646,189
Granted
Jul 26, 2011
Kind
B2
Abstract

The invention provides for a method for producing a 5-carbon alcohol in a genetically modified host cell. In one embodiment, the method comprises culturing a genetically modified host cell which expresses a first enzyme capable of catalyzing the dephosphorylation of an isopentenyl pyrophosphate (IPP) or dimethylallyl diphosphate (DMAPP), such as a Bacillus subtilis phosphatase (YhfR), under a suitable condition so that 5-carbon alcohol is 3-methyl-2-buten-1-ol and/or 3-methyl-3-buten-1-ol is produced. Optionally, the host cell may further comprise a second enzyme capable of reducing a 3-methyl-2-buten-1-ol to 3-methyl-butan-1-ol, such as a reductase.

Claims (21)

1. A method for producing a 5-carbon alcohol in a genetically modified host cell, the method comprising: culturing a genetically modified host cell under a suitable condition, wherein the genetically modified host cell is transformed with a first nucleic acid construct encoding a first enzyme capable of catalyzing the dephosphorylation of an isopentenyl pyrophosphate (IPP) or dimethylallyl diphosphate (DMAPP), such that the culturing results in the genetically modified host cell producing a 5-carbon alcohol, wherein the 5-carbon alcohol is 3-methyl-2-buten-1-ol, or 3-methyl-3-buten-1-ol.

2. The method of claim 1 , further comprising the step of: (b) recovering the produced 5-carbon alcohol, wherein the recovering step is concurrent or subsequent to the culturing step.

3. The method of claim 1 , wherein the first enzyme is Bacillus sp. YhfR , having the amino acid sequence of SEQ ID NO:70.

4. The method of claim 1 , wherein the genetically modified host cell is further transformed with a second nucleic acid construct encoding a second enzyme capable of reducing a 3-methyl-2-buten-1-ol, such that the culturing results in the genetically modified host cell producing 3-methyl-butan-1-ol, wherein the first nucleic acid construct and the second nucleic acid construct reside on the same nucleic acid or two separate nucleic acids.

5. The method of claim 4 , wherein the second enzyme is a reductase.

6. The method of claim 5 , wherein the reductase is an Escherichia coli, Saccharomyces cerevisiae, Torulaspora delbrueckii , or Kluyveromyces lactis fatty acid synthase.

7. The method of claim 5 , wherein the reductase is an OYE family member.

8. The method of claim 7 , wherein the reductase is a Saccharomyces sp. OYE family member.

9. The method of claim 7 , wherein the OYE family member is OYE2.

10. The method of claim 7 , wherein the OYE family member is OYE3.

11. The method of claim 4 , further comprising the step of: (b) recovering the produced 3-methyl-butan-1-ol, wherein the recovering step is concurrent or subsequent to the culturing step.

12. The method of claim 1 , wherein the 5-carbon alcohol is 3-methyl-2-buten-1-ol.

13. The method of claim 1 , wherein the 5-carbon alcohol is 3-methyl-3-buten-1-ol.

14. The method of claim 1 , wherein the host cell is a eubacteria.

15. The method of claim 14 , wherein the host cell is one selected from the Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsielia, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla, Synechococcus, Synechocystis , and Paracoccus taxonomical classes.

16. The method of claim 14 , wherein the host cell is Escherichia coli.

17. The method of claim 1 , wherein the host cell is an algal, fungal, insect, or an isolated mammalian cell line.

18. The method of claim 17 , wherein the host cell is a yeast.

19. The method of claim 18 , wherein the host cell is Saccharomyces cerevisae.

20. A genetically modified host cell transformed with a first nucleic acid construct encoding a first enzyme capable of catalyzing the dephosphorylation of an isopentenyl pyrophosphate (IPP) or dimethylallyl diphosphate (DMAPP), which under a suitable condition produces 3-methyl-2-buten-1-ol, or 3-methyl-3-buten-1-ol, wherein said host cell prior to genetic modification does not produce 3-methyl-2-buten-1-ol, 3-methyl-3-buten-1-ol and 3-methyl-butan-1-ol.

21. The genetically modified host cell of claim 20 , further comprising a second nucleic acid construct encoding a second enzyme capable of reducing a 3-methyl-2-buten-1-ol, wherein the first nucleic acid construct and the second nucleic acid construct reside on the same nucleic acid or two separate nucleic acids.

Assignments (2)
CONFIRMATORY LICENSE Recorded Feb 22, 2010
From: REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE
To: ENERGY, UNITED STATES DEPARTMENT OF
Reel/Frame 023972/0372 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 16, 2010
From: CHOU, HOWARD H.; KEASLING, JAY D.
To: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Reel/Frame 023943/0047 →
Continuity (2)
Provisional Application 60947285 · Jun 29, 2007
Related Publication 20100205855A1 · Aug 19, 2010