Light-powered, biological methyl laurate production from CO2 and water
AssigneeArizona Board of Regents on behalf of Arizona State University
InventorWillem Vermaas
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Abstract
The disclosure relates to the production of methyl laurate by genetically engineered photosynthetic microorganisms. In particular, provided herein are methods and compositions for producing methyl laurate from carbon dioxide and water in genetically engineered cyanobacteria and other photosynthetic microorganisms.
Claims12
A genetically engineered Synechocystis sp. PCC 6803 cyanobacterium, wherein the engineered cyanobacterium has been transformed with one or more plasmid vectors comprising (i) an expression cassette comprising a first nucleotide sequence encoding a Mycobacterium marinum fatty acid methyl transferase (FAMT) and (ii) a second nucleotide sequence encoding a thioesterase, wherein the genetically engineered cyanobacterium includes an inactivated nitrogen regulatory protein P-II gene, g1nB, and wherein the genetically engineered cyanobacterium has an increased production of methyl laurate as compared to methyl laurate production by a native Synechocystis sp. PCC 6803 cyanobacterium.
A method of producing methyl laurate with a genetically modified cyanobacterium, the method comprising:
- (a) introducing an expression cassette comprising a nucleic acid encoding a Mycobacterium marinum fatty acid methyl transferase (FAMT) into a cyanobacterium to obtain a genetically modified cyanobacterium comprising the expression cassette, wherein the genetically modified cyanobacterium further comprises an exogenous nucleotide sequence encoding a thioesterase, wherein the genetically engineered cyanobacterium includes an inactivated nitrogen regulatory protein P-II gene, g1nB, and wherein the genetically engineered cyanobacterium has an increased production of methyl laurate as compared to methyl laurate production by a native Synechocystis sp. PCC 6803 cyanobacterium,
- (b) culturing the genetically modified cyanobacterium in a culture under conditions in which the nucleic acid encoding the FAMT is expressed and the genetically modified cyanobacterium produces and excretes methyl laurate;
- (c) contacting the culture with a dodecane layer, whereby methyl laurate collects in the dodecane layer;
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- (d) isolating methyl laurate from the dodecane layer.
A method of increasing fatty acid or fatty acyl ester production of a genetically modified strain of cyanobacterium, the method comprising:
- (a) obtaining a genetically modified cyanobacterium, wherein the genetically modified cyanobacterium includes an inactivated nitrogen regulatory protein P-II gene g1nB;
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- (b) introducing into the genetically modified cyanobacterium a first nucleotide sequence encoding a FAMT and a second nucleotide sequence encoding a fatty acid Acyl-ACP thioesterase to produce a resulting genetically modified cyanobacterium exhibiting increased fatty acid or fatty acyl ester production relative to fatty acid or fatty acyl ester production of the genetically modified cyanobacterium which does not comprise the first nucleotide sequence.
A genetically modified Synechocystis cyanobacterium strain comprising a first heterologous gene encoding a fatty acid methyl transferase (FAMT) and a second heterologous gene encoding a thioesterase, whereas the genetically modified Synechocystis cyanobacterium strain is capable of increased production of methyl laurate compared to an unmodified Synechocystis cyanobacterium strain not comprising the first heterologous gene, wherein the genetically modified Synechocystis cyanobacterium includes an inactivated nitrogen regulatory protein P-II gene, g1nB.
Dependencies are inferred from the claim text.