Increase in activity for the C-terminal Pro-X bond by site-directed mutagenesis of Gly137 to Ala in carboxypeptidase Z

K Ratanakhanokchai; B R Lee; Y Kobayashi; M Takeuchi

doi:10.1271/bbb.60.496

Increase in activity for the C-terminal Pro-X bond by site-directed mutagenesis of Gly137 to Ala in carboxypeptidase Z

Biosci Biotechnol Biochem. 1996 Mar;60(3):496-7. doi: 10.1271/bbb.60.496.

Authors

K Ratanakhanokchai¹, B R Lee, Y Kobayashi, M Takeuchi

Affiliation

¹ Laboratory of Molecular Biology and Microbial Chemistry, Tokyo University of Agriculture and Technology, Japan.

PMID: 8901112
DOI: 10.1271/bbb.60.496

Abstract

A mutant carboxypeptidase Z from Absidia zychae in which Gly137 was replaced by Ala by site-directed mutagenesis was constructed and expressed in Saccharomyces cerevisiae YPH250. The mutant enzyme hydrolyzed C-terminal Pro-X bonds (X = amino acid) more efficiently than the wild-type enzyme and sequentially released amino acids from the C-termini of oligopeptides.

MeSH terms

Alanine / metabolism
Amino Acid Sequence
Angiotensin II / metabolism
Base Sequence
Binding Sites
Bradykinin / metabolism
Carboxypeptidases / genetics*
Carboxypeptidases / metabolism*
Glycine / metabolism
Kinetics
Molecular Sequence Data
Mutagenesis, Site-Directed*
Substrate Specificity

Substances

Angiotensin II
Carboxypeptidases
serine carboxypeptidase
carboxypeptidase Z
Alanine
Bradykinin
Glycine