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AssigneeIowa State University Research Foundation, Inc.
InventorsShivani Garg, Huanan Jin, Marna Yandeau-Nelson, Basil J. Nikolau
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A mutant Bacillus subtilis, which does not express a functional KASIIIA and/or KASIIIB, and method of making; a mutant Rhodospirillum rubrum, which does not express a functional PhaC1, PhaC2, and/or PhaC3, and method of making; method of characterizing substrate specificity of KASIII; method of making mutant KASIII with altered substrate specificity and/or altered level of activity and nucleic acid, vector, host cell/organism, and mutant KASIII; an in vitro, high-throughput spectrophotometric method of assaying KASIII activity; and materials and methods for using KASIII for production of bi-functional fatty acids and the materials so produced.
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.
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