Screening and isolation of cephalosporin acylase enzyme-producing bacteria and optimization of enzyme production process

Abstract

Objectives: Cephalosporin acylases are highly specialized peptidases, which are capable for cleaving the amide bond between a cephalosporin nucleus and its side chain. This enzyme is used in the enzymatic conversion of cephalosporin C to the cephem nucleus (7-ACA). 7-ACA is a key intermediate for manufacturing of semi synthetic cephalosporins in the pharmaceutical industries. Thus, achieving an efficient, safe and economical production routes is of great economical and bio-environmental importance. Methods: The aim of this study was to screen soil bacteria for identification of cephalosporin acylase producing pseudomonas strains with spectrophotometric method. In spectrophotometric method, the enzyme activity, production of 7-ACA and formation of imine group with ?DAB are determined by a colorimetric reaction. After isolation and identification of enzyme producing strain, the effect of incubation time, temperature, culture medium pH and glutaric acid concentration on production of enzyme were studied. Results: In this research, out of 215 soil-isolated strains, only one strain showed noticeable cephalosporin acylase activity. Selected strain showed optimized activity by incubation at 35ط¢آ° C for 48 hours in a pH=8 and in the presence of 2 gram/liter glutaric acid. Conclusion: Findings of this study indicates that spectrophotometric method is an effective and rapid method for identification and isolation of cephalosporin acylase producing microorganisms.