IP Library Granted Patent US 8,445,257
Granted Patent B2
US 8,445,257 · App. 13/338,579 · Granted May 21, 2013

Production of branched-chain alcohols by photosynthetic microorganisms

Inventors: Paul Gordon Roessler (San Diego, CA); Bo Liu (San Diego, CA); Jessica Roxane Kristof (San Diego, CA)
Assignee: Synthetic Genomics, Inc.
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Quick Facts
Patent No.
US 8,445,257
App. No.
13/338,579
Granted
May 21, 2013
Kind
B2
Abstract

The present invention provides genes, polypeptides and expression constructs therefor, recombinant photosynthetic microorganisms, and method of use thereof, such as for the production of branched-chain alcohols (including 2-methyl-1-butanol, 3-methyl-1-butanol, and isobutanol) and derivatives thereof for a variety of uses.

Claims (20)

1. A recombinant photosynthetic microorganism comprising at least one heterologous nucleic acid sequence that encodes a branched-chain 2-ketoacid decarboxylase, wherein the recombinant photosynthetic microorganism produces a branched-chain aldehyde.

2. The recombinant photosynthetic microorganism of claim 1 , wherein the branched-chain 2-ketoacid decarboxylase is selected from the group consisting of a Saccharomyces cerevisiae PDC 1 gene product, a Pichia stipitis PDC 1 gene product, a Saccharomyces cerevisiae PDC5 gene product, a Saccharomyces cerevisiae PDC6 gene product, a Saccharomyces cerevisiae ARO10 gene product, a Saccharomyces cerevisiae THI3 gene product, a Pichia stipitis PDC 3-6 Kivd gene product, a Pichia stipitis PDC2 gene product, a Mycobacterium tuberculosis KDC gene product, a Lactococcus lactis KDCa gene product, and a variant or homolog of any thereof.

3. The recombinant photosynthetic microorganism of claim 2 , wherein the heterologous nucleic acid sequence encodes a Lactococcus lactis KDCa gene product or a variant thereof.

4. The recombinant photosynthetic microorganism of claim 3 , wherein the heterologous nucleic acid sequence encodes a Lactococcus lactis KDCa-S286Y, KDCa S286U, or F381W gene product, or a variant of any thereof.

5. The recombinant photosynthetic microorganism of claim 2 , wherein the heterologous nucleic acid sequence encodes a Pichia stipitis PDC3-6 gene product or a variant thereof.

6. The recombinant photosynthetic microorganism of claim 1 , wherein the heterologous nucleic acid sequence that encodes a branched-chain 2-ketoacid decarboxylase is cloned in a vector designed for integration of the heterologous nucleic acid sequence into the host genome.

7. The recombinant photosynthetic microorganism of claim 1 , wherein the heterologous nucleic acid sequence that encodes a branched-chain 2-ketoacid decarboxylase is operably linked to an inducible promoter.

8. The recombinant photosynthetic microorganism of claim 2 , wherein said at least one heterologous nucleic acid sequence further encodes one or more gene products selected from the group consisting of an acetolactate synthase gene product, a ketol-acid reductoisomerase gene product, and a dihydroxyacid dehydratase gene product.

9. The recombinant photosynthetic microorganism of claim 8 , wherein said one or more gene products is an acetolactate synthase gene product.

10. The recombinant photosynthetic microorganism of claim 2 , wherein said at least one heterologous nucleic acid sequence further encodes one or more gene products selected from the group consisting of a 2-isopropylmalate synthase gene product, a 3-isopropylmalate dehydratase gene product, and a 3-isopropylmalate dehydrogenase gene product.

11. The recombinant photosynthetic microorganism of claim 10 , wherein said one or more gene products is a 2-isopropylmalate synthase gene product.

12. The recombinant photosynthetic microorganism of claim 2 , wherein said at least one heterologous nucleic acid sequence further encodes one or more gene products selected from the group consisting of a homoserine dehydrogenase gene product, a homoserine kinase gene product, a threonine synthase gene product, and a threonine ammonia-lyase gene product.

13. The recombinant photosynthetic microorganism of claim 12 , wherein said at least one heterologous nucleic acid sequence further encodes a threonine ammonia-lyase gene product.

14. The recombinant photosynthetic microorganism of claim 1 , wherein the photosynthetic microorganism is a eukaryotic microalga.

15. The recombinant photosynthetic microorganism of claim 13 , wherein the photosynthetic microorganism is selected from the group consisting of Achnanthes, Amphiprora, Amphora, Ankistrodesmus, Asteromonas, Boekelovia, Borodinella, Botryococcus, Bracteococcus, Chaetoceros, Carteria, Chlamydomonas, Chlorococcum, Chlorogonium, Chlorella, Chroomonas, Chrysosphaera, Cricosphaera, Crypthecodinium, Cryptomonas, Cyclotella, Dunaliella, Ellipsoidon, Emiliania, Eremosphaera, Ernodesmius, Euglena, Franceia, Fragilaria, Gloeothamnion, Haematococcus, Halocafeteria, Hymenomonas, Isochrysis, Lepocinclis, Micractinium, Monoraphidium, Nannochloris, Nannochloropsis, Navicula, Neochloris, Nephrochloris, Nephroselmis, Nitzschia, Ochromonas, Oedogonium, Oocystis, Ostreococcus, Pavlova, Parachlorella, Pascheria, Phaeodactylum, Phagus, Platymonas, Pleurochrysis, Pleurococcus, Prototheca, Pseudochlorella, Pyramimonas, Pyrobotrys, Scenedesmus, Skeletonema, Spyrogyra, Stichococcus, Tetraselmis, Thalassiosira, Viridiella, and Volvox.

16. The recombinant photosynthetic microorganism of claim 1 , wherein the photosynthetic microorganism is a cyanobacterium.

17. The recombinant photosynthetic microorganism of claim 16 , wherein the photosynthetic microorganism is selected from the group consisting of Chamaesiphon, Chroococcus, Cyanobacterium, Cyanobium, Dactylococcopsis, Gloeobacter, Gloeocapsa, Gloeothece, Microcystis, Prochlorococcus, Prochloron, Synechococcus, Synechocystis, Chroococcidiopsis, Cyanocystis, Dermocarpella, Myxosarcina, Pleurocapsis, Stanieria, Xenococcus, Arthrospira, Borzia, Crinalium, Geitlerinema, Halospirulina, Leptolyngbya, Limnothrix, Lyngbya, Microcoleus, Oscillatoria, Planktothrix, Prochlorothrix, Pseudanabaena, Spirulina, Starria, Symploca, Trichodesmium, and Tychonema, Anabaena, Anabaenopsis, Aphanizomenon, Calothrix, Cyanospira, Cylindrospermopsis, Cylindrospermum, Nodularia, Nostoc, Chlorogloeopsis, Fischerella, Geitleria, Nostochopsis, Iyengariella, Stigonema, Rivularia, Scytonema, and Tolypothri.

18. The recombinant photosynthetic microorganism of claim 17 , wherein the photosynthetic microorganism is Synechococcus or Synechocystis.

19. The recombinant photosynthetic microorganism of claim 1 , wherein the photosynthetic microorganism produces 2-methyl-1-propanal, 2-methyl-1-butanal, or 3-m ethyl-1-butanal.

20. The recombinant photosynthetic microorganism of claim 19 , wherein the photosynthetic microorganism produces 2-methyl-1-propanal.

Assignments (2)
RELEASE OF SECURITY INTEREST Recorded Nov 10, 2020
From: OXFORD FINANCE LLC
To: SYNTHETIC GENOMICS, INC.; GENOVIA BIO, LLC; GREEN RESOURCES, LLC; SGI-DNA, INC.; SYNTHETIC GENOMICS VACCINES, INC.
Reel/Frame 054372/0822 →
SECURITY INTEREST Recorded Mar 20, 2019
From: SYNTHETIC GENOMICS, INC.
To: OXFORD FINANCE LLC
Reel/Frame 048655/0406 →
Continuity (3)
Continuation 12635653 · Dec 10, 2009
Provisional Application 61121522 · Dec 10, 2008
Related Publication 20120094362A1 · Apr 19, 2012