A penicillin G acylase mutant constructed by means of gene engineering; when compared with wild-type penicillin G acylase derived from Achromobacter sp.CCM 4824, the synthesis performance of the present penicillin G acylase mutant is greatly improved, the synthesis product/hydrolysis product value S/H reaching a maximum of 22.3, 3.9 times that of the wild-type enzyme, being able to efficiently catalyse the synthesis of various β-lactam antibiotics. When the ratio of the side chain to the mother nucleus is 1.05:1, the conversion rate of the 6-APA, 7-ADCA, 7-ACCA, and 7-APRA of the mother nucleus is above 99.0%.
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