Production of hydroxylated fatty acids in
First Claim
1. A method for using sucrose to produce an oil comprising triacylglycerides that comprise ricinoleic acid, or a product produced from the oil, the method comprising:
- cultivating a cell of a recombinant oleaginous microalga of the species Prototheca moriformis, the cell comprising;
nucleic acids operable to enhance oleic acid production so as to elevate the substrate levels for an exogenous oleate 12-hydroxylase by expressing a product of an exogenous gene encoding an active stearoyl ACP desaturase and decreasing or eliminating the expression of an enzyme encoded by one or more endogenous genes that encode a fatty acid desaturase by directly targeting said endogenous gene(s) or RNA products of said gene(s); and
an exogenous nucleic acid operable to express the active oleate 12-hydroxylase, so as to synthesize the ricinoleic acid; and
an exogenous nucleic acid operable to express an active invertase; and
feeding sucrose to the cell so as to produce the oil comprising triacylglycerides comprising ricinoleic acid, wherein the ricinoleic acid is detectable by gas chromatography-mass spectrometry.
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Abstract
Methods and compositions for the production of oil, fuels, oleochemicals, and other compounds in recombinant microorganisms are provided, including oil-bearing microorganisms and methods of low cost cultivation of such microorganisms. Microalgal cells containing exogenous genes encoding, for example, a lipase, a sucrose transporter, a sucrose invertase, a fructokinase, a polysaccharide-degrading enzyme, a keto acyl-ACP synthase enzyme, a fatty acyl-ACP thioesterase, a fatty acyl-CoA/aldehyde reductase, a fatty acyl-CoA reductase, a fatty aldehyde reductase, a fatty acid hydroxylase, a desaturase enzyme, a fatty aldehyde decarbonylase, and/or an acyl carrier protein are useful in manufacturing transportation fuels such as renewable diesel, biodiesel, and renewable jet fuel, as well as oleochemicals such as functional fluids, surfactants, soaps and lubricants.
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Citations
7 Claims
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1. A method for using sucrose to produce an oil comprising triacylglycerides that comprise ricinoleic acid, or a product produced from the oil, the method comprising:
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cultivating a cell of a recombinant oleaginous microalga of the species Prototheca moriformis, the cell comprising; nucleic acids operable to enhance oleic acid production so as to elevate the substrate levels for an exogenous oleate 12-hydroxylase by expressing a product of an exogenous gene encoding an active stearoyl ACP desaturase and decreasing or eliminating the expression of an enzyme encoded by one or more endogenous genes that encode a fatty acid desaturase by directly targeting said endogenous gene(s) or RNA products of said gene(s); and an exogenous nucleic acid operable to express the active oleate 12-hydroxylase, so as to synthesize the ricinoleic acid; and
an exogenous nucleic acid operable to express an active invertase; andfeeding sucrose to the cell so as to produce the oil comprising triacylglycerides comprising ricinoleic acid, wherein the ricinoleic acid is detectable by gas chromatography-mass spectrometry. - View Dependent Claims (2, 4, 5)
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3. A microalga cell of the species Prototheca moriformis, the cell comprising:
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nucleic acids operable to enhance oleic acid production so as to elevate the substrate levels for an exogenous oleate 12-hydroxylase by expressing a product of an exogenous gene encoding an active stearoyl ACP desaturase and decreasing or eliminating the expression of an enzyme encoded by one or more endogenous genes that encode a fatty acid desaturase by directly targeting said endogenous gene(s) or RNA products of said gene(s);
an exogenous gene encoding the active oleate 12-hydroxylase; andan exogenous gene encoding an invertase so as to enable the cell to use sucrose to synthesize ricinoleic acid in an amount detectable by gas chromatography-mass spectrometry. - View Dependent Claims (6, 7)
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Specification