Production of enantiomerically pure (S)-β-phenylalanine and (R)-β-phenylalanine by penicillin G acylase from Escherichia coli in aqueous medium

Abstract

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.

Synthesis of Cephalexin in Aqueous Medium with Carrier-bound and Carrier-free Penicillin Acylase Biocatalysts

“…3.5.1.11) is a flexible enzyme that is able to catalyze several reactions of organic synthesis [1][2][3][4][5][6]. Among those reactions, the synthesis of semisynthetic penicillins and cephalosporins from the corresponding β-lactam nuclei and suitable acyl donors is of paramount importance to the pharmaceutical industry [7].…”

Hydrolysis of Amides

“…Most of these methods are chemical in nature [319][320][321], but enzymatic methods applying, for instance, a lipase [322,323], aminoacylase [324], penicillin G acylase [325], peptide deformylase [9], and aminomutase [326] have also been described (for a review see Reference [327]). An enzyme with activity toward a b-amino acid amide was described in 2005 by Komeda and Asano [82].…”

Quantifying the efficiency of Saccharomyces cerevisiae translocation tags

“…Enzyme localization to the S. cerevisiae peroxisome takes advantage of fatty acid degradation intermediates. Additionally, the peroxisome may provide a more suitable environment for enzymes with a high pH optimum, such as those in β‐lactam biosynthesis (Li, Cheng, Wei, Ren, & Zhang, ).…”

“…Additionally, the peroxisome may provide a more suitable environment for enzymes with a high pH optimum, such as those in β-lactam biosynthesis (Li, Cheng, Wei, Ren, & Zhang, 2007).…”