Gene construct encoding mutant thioesterase, mutant thioesterase encoded thereby, transformed host cell containing the gene construct, and method of using them to produce medium-chain fatty acids
AssigneesWisconsin Alumni Research Foundation, The Penn State Research Foundation
InventorsBrian F. Pfleger, Nestor Jose Hernandez-Lozada, Costas Maranas, Matthew Grisewood
Life of this patent
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Abstract
Unnatural, mutated thioesterases having an amino acid sequence that is at least 80% identical to SEQ ID NO:1 and having substitutions at one or more of amino acid positions I107, R108, L109, S122, M141, E142, Y145, and L146, gene constructs encoding and configured to express the mutated thioesterases in a transformed host cell, and host cells transformed to contain the gene constructs.
Claims20
An unnatural, mutated protein comprising an amino acid sequence that is at least 85% identical to SEQ ID NO:1 and comprises a substitution at a position aligning to I107 of SEQ ID NO:1, a position aligning to R108 of SEQ ID NO:1, a position aligning to S122 of SEQ ID NO:1, a position aligning to M141 of SEQ ID NO:1, a position aligning to Y145 of SEQ ID NO:1, a position aligning to L146 of SEQ ID NO:1, or a combination thereof, wherein:
- the protein comprises at least one of:
- a lysine at the position aligning to S122 of SEQ ID NO:1;
- a lysine at the position aligning to Y145 of SEQ ID NO:1;
- and
- a lysine at the position aligning to L146 of SEQ ID NO:1;
- and
- the protein has at least one of enhanced thioesterase activity and enhanced thioesterase specificity in catalyzing the hydrolysis of a medium-chain acyl-acyl carrier protein substrate or a medium-chain acyl-CoA substrate to yield a free fatty acid or a free fatty acid derivative compared to an unaltered protein of SEQ ID NO:1.
Dependencies are inferred from the claim text.