Methods of producing omega-hydroxylated fatty acid derivatives
AssigneeGenomatica, Inc.
InventorsAndreas W. Schirmer, Haibo Wang, Stephen B. Del Cardayre, Zhihao Hu, Louis G. Hom, Baolong Zhu, Cindy Chang, Emanuela E. Popova
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Abstract
The disclosure relates to omega-hydroxylated fatty acid derivatives and methods of producing them. Herein, the disclosure encompasses a novel and environmentally friendly production method that provides omega-hydroxylated fatty acid derivatives at high purity and yield. Further encompassed are recombinant microorganisms that produce omega-hydroxylated fatty acid derivatives through selective fermentation.
Claims22
A recombinant microorganism for producing an ω-hydroxy fatty acid derivative in vivo when grown in a fermentation broth in a presence of a carbon source from a renewable feedstock, said microorganism comprising a pathway engineered to express:
- (i) at least two nucleic acid sequences encoding a polypeptide comprising:
- (a) a thioesterase of EC 3.1.2.-, 3.1.1.5, or 3.1.2.14;
- or an ester synthase of EC 2.3.1.75 or EC 2.3.1.20;
- or a combination thereof;
- and
- (b) a modified ω-hydroxylase of EC 1.14.15.3;
- or
- (ii) at least three nucleic acid sequences encoding a polypeptide comprising:
- (a) an acyl-ACP reductase of EC 1.2.1.80 or EC 1.2.1.42;
- (b) an alcohol dehydrogenase of EC 1.1.-.-;
- and
- (c) a modified ω-hydroxylase of EC 1.14.15.3
- wherein the modified ω-hydroxylase comprises a CYP153A polypeptide having at least 90% sequence identity to SEQ ID NO:2;
- and
- wherein the modified ω-hydroxylase comprises the mutation G307A and is further modified to comprise at least one additional mutation at an amino acid position corresponding to one or more of positions 6, 11, 27, 40, 41, 82, 116, 129, 131, 132, 134, 136, 138, 141, 142, 144, 149, 153, 154, 157, 161, 168, 178, 205, 228, 231, 233, 258, 271, 304, 308, 309, 407, 415, and 419 of SEQ ID NO:2.
Dependencies are inferred from the claim text.