Methods of using ketoreductase polypeptides for reduction of acetophenones

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. A method for stereoselectively reducing 2,6-dichloro-3′-fluoroacetophenone substrate, optionally substituted at one or more positions selected from the group consisting of 4′ and 5′, to the corresponding substituted (S)-1-phenethanol, which comprises contacting the substrate with an engineered ketoreductase under reaction conditions suitable for stereoselectively reducing or converting the substrate to the corresponding substituted (S)-1-phenethanol product, wherein the engineered ketoreductase is derived from a wild-type Lactobacillus ketoreductase and wherein said engineered ketoreductase comprises an amino acid sequence at least 90% identical to the reference sequence of SEQ ID NO: 2, 4, or 98; and wherein said engineered ketoreductase is capable of stereoselectively reducing 2,6-dichloro-3′-fluoroacetophenone to (S)-1-(2,6-dichloro-3-fluorophenyl) ethanol.