Mutant thioesterases
Mutant thioesterases having enhanced medium chain substrate activity, polynucleotides encoding and configured to express the mutant thioesterases in a transformed host cell, host cells transformed to contain the polynucleotides, and methods of using same.
1. An unnatural, mutated protein comprising an amino acid sequence at least 80% identical to SEQ ID NO:4, wherein:
the amino acid sequence comprises:
a serine at a position corresponding to position 59 of SEQ ID NO:4; and
an arginine at a position corresponding to position 296 of SEQ ID NO:4; and
the protein exhibits thioesterase activity.
2. An unnatural, mutated protein comprising an amino acid sequence at least 85% identical to SEQ ID NO:4, wherein:
the amino acid sequence comprises:
a residue other than alanine at a position corresponding to position 59 of SEQ ID NO:4; and
a residue other than lysine at a position corresponding to position 296 of SEQ ID NO:4; and
the protein exhibits thioesterase activity.
3. The protein of claim 2 , wherein the amino acid sequence comprises one or more of:
a serine, cysteine, threonine, asparagine, tyrosine, or glutamine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine, lysine, or histidine at the position corresponding to position 296 of SEQ ID NO:4.
4. The protein of claim 2 , wherein the amino acid sequence comprises one or more of:
a serine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine at the position corresponding to position 296 of SEQ ID NO:4.
5. The protein of claim 2 , wherein the amino acid sequence is at least 90% identical to SEQ ID NO:4.
6. The protein of claim 5 , wherein the amino acid sequence comprises one or more of:
a serine, cysteine, threonine, asparagine, tyrosine, or glutamine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine, lysine, or histidine at the position corresponding to position 296 of SEQ ID NO:4.
7. The protein of claim 5 , wherein the amino acid sequence comprises one or more of:
a serine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine at the position corresponding to position 296 of SEQ ID NO:4.
8. The protein of claim 2 , wherein the amino acid sequence is at least 95% identical to SEQ ID NO:4.
9. The protein of claim 8 , wherein the amino acid sequence comprises one or more of:
a serine, cysteine, threonine, asparagine, tyrosine, or glutamine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine, lysine, or histidine at the position corresponding to position 296 of SEQ ID NO:4.
10. The protein of claim 8 , wherein the amino acid sequence comprises one or more of:
a serine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine at the position corresponding to position 296 of SEQ ID NO:4.
11. The protein of claim 8 , wherein the amino acid sequence comprises:
a serine, cysteine, threonine, asparagine, tyrosine, or glutamine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine, lysine, or histidine at the position corresponding to position 296 of SEQ ID NO:4.
12. The protein of claim 8 , wherein the amino acid sequence comprises:
a serine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine at the position corresponding to position 296 of SEQ ID NO:4.
13. A method of producing a fatty acid derivative comprising contacting the unnatural, mutated protein of claim 2 with an acyl-acyl carrier protein substrate or an acyl-coenzyme A substrate under conditions that permit production of the fatty acid derivative.
14. The method of claim 13 , wherein the fatty acid derivative comprises a C8 fatty acid derivative.
15. The method of claim 14 , wherein the C8 fatty acid derivative comprises octanoic acid.
16. The method of claim 13 , wherein the amino acid sequence is at least 95% identical to SEQ ID NO:4.
17. The method of claim 16 , wherein the amino acid sequence comprises:
a serine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine at the position corresponding to position 296 of SEQ ID NO:4.
18. The method of claim 14 , wherein the amino acid sequence is at least 95% identical to SEQ ID NO:4 and comprises:
a serine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine at the position corresponding to position 296 of SEQ ID NO:4.
19. The method of claim 15 , wherein the amino acid sequence is at least 95% identical to SEQ ID NO:4 and comprises:
a serine at the position corresponding to position 59 of SEQ ID NO:4; and
an arginine at the position corresponding to position 296 of SEQ ID NO:4.