Abstract: An objective of the present invention is to provide efficient methods for producing (R)-2-chloromandelamide with high optical purity. Another objective of the present invention is to provide novel methods for producing ?-ketoamide reductases that reduce 2-chlorobenzoyl formamide to (R)-2-chloromandelamide with high optical purity, using NADPH as the coenzyme. An enzyme exhibiting high stereoselectivity was purified from a number of Saccharomyces cerevisiae enzymes with 2-chlorobenzoyl formamide-reducing activity, and the biochemical properties of the purified enzyme were analyzed. The analysis of a partial internal amino acid sequence of the purified enzyme revealed that the enzyme may be encoded by the putative open reading frame (ORF) YDL124w reported in the genome analysis. YDL124w was cloned and expressed in E. coli, and was subsequently shown to encode the ?-ketoamide reductase.

Abstract: An objective of the present invention is to provide efficient methods for producing (R)-2-chloromandelamide with high optical purity. Another objective of the present invention is to provide novel methods for producing ?-ketoamide reductases that reduce 2-chlorobenzoyl formamide to (R)-2-chloromandelamide with high optical purity, using NADPH as the coenzyme. An enzyme exhibiting high stereoselectivity was purified from a number of Saccharomyces cerevisiae enzymes with 2-chlorobenzoyl formamide-reducing activity, and the biochemical properties of the purified enzyme were analyzed. The analysis of a partial internal amino acid sequence of the purified enzyme revealed that the enzyme may be encoded by the putative open reading frame (ORF) YDL124w reported in the genome analysis. YDL124w was cloned and expressed in E. coli, and was subsequently shown to encode the ?-ketoamide reductase.