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US 12,416,028 B2Granted
Biosynthetic production of gamma- and delta-lactones using cytochrome P450 enzymes having subterminal hydroxylase activity
AssigneeConagen Inc.
InventorsHui Chen, Xiaodan Oliver Yu
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Abstract
Provided herein are methods for making lactones comprising incubating a cellular system expressing a heterologous cytochrome P450 (CYP450) protein with a medium comprising straight chain fatty acids to produce subterminal hydroxy fatty acids, and incubating a yeast cell culture with the subterminal hydroxy fatty acids to produce lactones.
Claims17
A method of making lactones, the method comprising:
- (a) incubating a cellular system expressing a heterologous cytochrome P450 (CYP450) protein with a medium comprising a straight chain fatty acid to produce a subterminal hydroxy fatty acid, wherein the straight chain fatty acid and the subterminal hydroxy fatty acid have 5 to 40 carbon atoms, and wherein the heterologous cytochrome P450 protein comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of a cytochrome P450 protein from Umbelopsis isabellina, and either:
- (b1) incubating the subterminal hydroxy fatty acid with a yeast cell culture to produce lactones, wherein the lactones comprise at least one gamma-lactone, at least one delta-lactone, or combinations thereof, or
- (b2) incubating the subterminal hydroxy fatty acids in an acidic environment to produce lactones, wherein the lactones comprise at least one gamma-lactone, at least one delta-lactone, or combinations thereof.
A method of making lactones, the method comprising:
- (a) incubating a cellular system expressing a heterologous cytochrome P450 protein with a medium comprising a straight chain fatty acid to produce a subterminal hydroxy fatty acid, wherein the heterologous cytochrome P450 protein comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of a cytochrome P450 protein from Umbelopsis isabellina;
- and
- wherein the straight chain fatty acid and the subterminal hydroxy fatty acid have 5 to 40 carbon atoms;
- (b) incubating the subterminal hydroxy fatty acid from step (a) with a cell culture to produce subterminal hydroxy fatty acid with the length shorter than the subterminal hydroxy fatty acid from step (a);
- and
- (c) incubating the subterminal hydroxy fatty acids obtained at the end of step (b) in an acidic environment to produce lactones, wherein the lactones comprise at least one gamma-lactone, at least one delta-lactone, or combinations thereof.
3 more elements
Dependencies are inferred from the claim text.