US 9,809,804 B2
Materials and methods for characterizing and using KASIII for production of bi-functional fatty acids
Shivani Garg, Ames, IA (US); Huanan Jin, Wuhan (CN); Marna Yandeau-Nelson, Ames, IA (US); and Basil J. Nikolau, Ames, IA (US)
Assigned to IOWA STATE UNIVERSITY RESEARCH FOUNDATION, INC., Ames, IA (US)
Appl. No. 14/762,791
Filed by IOWA STATE UNIVERSITY RESEARCH FOUNDATION, INC., Ames, IA (US)
PCT Filed Jan. 22, 2014, PCT No. PCT/US2014/012616
§ 371(c)(1), (2) Date Jul. 29, 2015,
PCT Pub. No. WO2014/116754, PCT Pub. Date Jul. 31, 2014.
Claims priority of provisional application 61/755,946, filed on Jan. 23, 2013.
Prior Publication US 2016/0090576 A1, Mar. 31, 2016
Int. Cl. C12N 1/20 (2006.01); C12N 1/12 (2006.01); C12P 7/64 (2006.01); C12N 9/10 (2006.01); C12Q 1/48 (2006.01); C12N 15/00 (2006.01); C12N 15/74 (2006.01); C07H 21/04 (2006.01)
CPC C12N 9/1029 (2013.01) [C12P 7/6409 (2013.01); C12Q 1/48 (2013.01); C12Y 203/0118 (2013.01); G01N 2333/91057 (2013.01)] 8 Claims
 
1. A method of characterizing substrate specificity of a 3-ketoacyl-acyl carrier protein (ACP) synthase III (KASIII), which method comprises expressing the KASIII, which is not expressed in wild-type Bacillus subtilis, in a mutant B. subtilis, which does not express a functional KASIIIA and does not express a functional KASIIIB, and wherein the KASIIIB of SEQ ID NO:66 comprises a mutation of Trp221 and a mutation of Val226, and assessing the production of co-functionalized fatty acids, whereupon the substrate specificity of a KASIII is characterized.