[Effect of mutagenesis at Ser 177 residue in penicillin G acylase on activity of the enzyme].
The technique of cassette and site-specific mutagenesis were used to study the role of residue No. 177 in penicillin G acylase (PGA, EC 3.5.1.11). Ser is conserved at residue No. 177 in all penicillin binding proteins. We got a series of mutants in which the amino acid at residue No. 177 was replaced by other amino acids through the site-specific and cassette mutagenesis, and we characterized the mutants by colony hybridization, NIPAB paper test and DNA sequence analysis. These mutants all show no activity of enzyme, even if the Ser residue was replaced by Thr, Gly and Ala respectively. The results show that Ser residue may be essential for substrate-binding or catalysis of PGA.
Wang
M
M
Shanghai Institute of Cell Biology, Academia Sinica.
Fei
J A
JA
Guo
L H
LH
Zhang
Q J
QJ
D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D010399:Penicillin Amidase / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D012694:Serine / Q000235:genetics*
D012694:Serine; D010399:Penicillin Amidase