Carboxyesterase biocatalysts
AssigneeGlaxoSmithKline Intellectual Property Development Limited
InventorsKristin K. Brown, Brent M. Dorr, Douglas E. Fuerst, Katherine Joyce Honicker, Lydia Sanchez Jordan, James Patrick Morrison, Nikolay V. Plotnikov, Markus Schober, Rama Voladri
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Abstract
The present disclosure provides engineered carboxyesterase enzymes that have the ability to catalyze amide bond formation. Also provided are polynucleotides encoding the carboxyesterase enzymes, host cells capable of expressing the engineered carboxyesterase enzymes, and methods of using the engineered carboxyesterase enzymes to make commercially valuable amides. Also provided are amides that are made using the engineered carboxyesterase enzymes.
Claims18
A carboxyesterase polypeptide having carboxyesterase activity and comprising an amino acid sequence that is at least 93%, or more identical to the amino acid sequence set forth in SEQ ID NO:
- 4, wherein the residue corresponding to X198 in SEQ ID NO:
- 4 is a leucine (L) residue.
A process for preparing an amide, wherein components are combined containing:
- (a) an ester substrate of the form R1-COOR2, wherein R1 is chosen from:
- an sp3 carbon with 0 to 3 alkyl substituents;
- and an aromatic ring, and R2 is chosen from:
- a methyl group;
- an ethyl group;
- and 1-6 carbon alkyl chains;
- (b) an amine substrate;
- (c) a carboxyesterase polypeptide comprising an amino acid sequence that is at least 93% identical to the amino acid sequence set forth in SEQ ID NO:
- 4, wherein the residue corresponding to X198 in SEQ ID NO:
- 4 is a L residue;
- and
- (d) a solvent.
Dependencies are inferred from the claim text.