Chemoenzymatic enantioselective synthesis of phenylglycine and phenylglycine amide by direct coupling of the Strecker synthesis with a nitrilase reaction

The conversion of rac -phenylglycinonitrile by different variants the nitrilase from Pseudomonas fluorescens EBC191 (EC 3.5.5.1) was studied and amounts chiral composition formed phenylglycine amide compared. Muteins that converted to extraordinarily high or were tested for chemoenzymatic enantioselective one-pot synthesis ( R )- S )-phenylglycine amide. combined initial step in traditional chemical Strecker which results formation benzaldehyde, cyanide, ammonia with enzymatic nitrile variants. aminonitrile optimized order obtain conditions allowed under mildly alkaline (pH 9.5) maximal yields phenylglycinonitrile in-situ racemization compound. racemic directly without any interposed purification cells Escherichia coli overexpressing recombinant application a mutant E. defect hydrolysing peptidase JM109? pepA ) expressing highly reaction- )-specific variant ee -values ? 95% up 81% relation initially added benzaldehyde. These indicated dynamic kinetic resolution involved )-phenylglycinonitrile used concurrent hydrolysis )-phenylglycine. addition resting synthesizing an forming final product and/or using strains intact aminopeptidase gene resulted preferred amide,