Ketoreductase polypeptides for the production of azetidinone

摘要

The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds.

主权项

1. An engineered polynucleotide encoding a ketoreductase polypeptide capable of converting the substrate, methyl-2-benzamidomethyl-3-oxobutyrate, to the product, 2S, 3R-methyl-2-benzamidomethyl-3-hydroxybutyrate, with a percent stereomeric excess of at least about 60%, wherein the encoded ketoreductase polypeptide comprises an amino acid sequence that is at least 85% identical to SEQ ID NO: 86, wherein the engineered polynucleotide encodes a ketoreductase polypeptide amino acid sequence comprising the following features: the residue corresponding to X94 is threonine; the residue corresponding to X199 is a histidine, and the residue corresponding to X202 is valine or leucine.