Production of enantiomerically pure (S)-beta-phenylalanine and (R)-beta-phenylalanine by penicillin G acylase from Escherichia coli in aqueous medium.
A new approach has been developed for the production of enantiomerically pure (S)-beta-phenylalanine (S-BPA) and (R)-beta-phenylalanine in aqueous medium based on enantioselective acylation and hydrolysis properties of penicillin G acylase from Escherichia coli. The acylation reaction was highly preferential for the acylation of (R)-BPA to form N-phenylacetyl-(R)-BPA using phenylacetamide as an acyl donor, which was separated and then hydrolyzed to (R)-BPA by the same enzyme at pH 7.5. The optimal acylation reaction was at pH 10, 25 degrees C with a 2:1 molar ratio of phenylacetamide to BPA, 8 IU ml(-1) enzyme and 150 mM BPA. These resulted in a conversion of about 50% BPA; enantiomeric excess of (S)-BPA and (R)-BPA separated were 98 and 99%, respectively.
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, PR China.
Dengchao
D
Li
Shiwei
S
Cheng
Dongzhi
D
Wei
Yuhong
Y
Ren
Derong
D
Zhang
D000083:Acetanilides / Q000378:metabolism; D000215:Acylation; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D010399:Penicillin Amidase / Q000378:metabolism*; D010649:Phenylalanine / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D013237:Stereoisomerism; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
D000083:Acetanilides; D010649:Phenylalanine; D010399:Penicillin Amidase