A single amino acid substitution converts gamma-glutamyltranspeptidase to a class IV cephalosporin acylase (glutaryl-7-aminocephalosporanic acid acylase).
The aspartyl residue at position 433 of gamma-glutamyltranspeptidase of Escherichia coli K-12 was replaced by an asparaginyl residue. This substitution enabled gamma-glutamyltranspeptidase to deacylate glutaryl-7-aminocephalosporanic acid, producing 7-aminocephalosporanic acid, which is a starting material for the synthesis of semisynthetic cephalosporins.
Suzuki
Hideyuki
H
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Oiwake-cho, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan. hideyuki@lif.kyoto-u.ac.jp.
Miwa
Chinatsu
C
Ishihara
Sayaka
S
Kumagai
Hidehiko
H
D019943:Amino Acid Substitution; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D048168:Escherichia coli K12 / Q000201:enzymology / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D016297:Mutagenesis, Site-Directed; D010399:Penicillin Amidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D013379:Substrate Specificity; D005723:gamma-Glutamyltransferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D004269:DNA, Bacterial; D005723:gamma-Glutamyltransferase; D010399:Penicillin Amidase; C046586:glutarylamidocephalosporanic acid acylase