	US 11,981,952 B2
	Methods of producing omega-hydroxylated fatty acid derivatives
Andreas W. Schirmer, San Diego, CA (US); Haibo Wang, San Diego, CA (US); Stephen B. Del Cardayre, San Diego, CA (US); Zhihao Hu, San Diego, CA (US); Louis G. Hom, San Diego, CA (US); Baolong Zhu, San Diego, CA (US); Cindy Chang, San Diego, CA (US); and Emanuela E. Popova, San Diego, CA (US)
Assigned to Genomatica, Inc., San Diego, CA (US)
Filed by Genomatica, Inc., San Diego, CA (US)
Filed on Jun. 29, 2020, as Appl. No. 16/915,454.
Application 16/915,454 is a continuation of application No. 14/897,285, granted, now 10,711,288, previously published as PCT/US2014/042594, filed on Jun. 16, 2014.
Claims priority of provisional application 61/835,464, filed on Jun. 14, 2013.
Prior Publication US 2021/0071212 A1, Mar. 11, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 7/6409 (2022.01); C12N 1/21 (2006.01); C12N 9/02 (2006.01); C12N 9/04 (2006.01); C12N 15/52 (2006.01); C12N 15/62 (2006.01); C12P 7/18 (2006.01); C12P 7/42 (2006.01); C12P 7/6436 (2022.01)

CPC C12P 7/6409 (2013.01) [C12N 9/0006 (2013.01); C12N 9/0008 (2013.01); C12N 9/0071 (2013.01); C12N 9/0077 (2013.01); C12N 15/52 (2013.01); C12N 15/62 (2013.01); C12P 7/18 (2013.01); C12P 7/42 (2013.01); C12P 7/6436 (2013.01); C12Y 114/14001 (2013.01); C12Y 114/15003 (2013.01); C07K 2319/00 (2013.01); C12Y 101/00 (2013.01); C12Y 102/01042 (2013.01); Y02E 50/10 (2013.01)]

22 Claims

1. 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

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.