Open AccessModeling and Experimental Analysis of Cephalosporin C Acylase and Its Mutant
Author(s) -
Yu Ren
Publication year - 2013
Publication title -
the open biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.308
H-Index - 11
ISSN - 1874-0707
DOI - 10.2174/1874070701307010030
Subject(s) - cephalosporin c , chemistry , cephalosporin antibiotic , mutant , cephalosporin , homology modeling , stereochemistry , biocatalysis , biochemistry , combinatorial chemistry , enzyme , d amino acid oxidase , amino acid , oxidase test , catalysis , antibiotics , gene , reaction mechanism
7-amino cephalosporanic acid (7-ACA) is the crucial intermediate for the synthesis of semi-synthetic antibiotics,which is currently prepared by two-step biocatalysis using D-amino acid oxidase and glutaryl-7-amino cephalosporanicacid acylase (GL-7-ACA acylase) starting from cephalosporin C (CPC). Compared with the two-step enzymaticmethod, one-step method is more efficient and economical. But, the available Cephalosporin C acylase (CPC acylase) alwaystake glutaryl-7-amino cephalosporanic acid (GL-7-ACA) as their primary substrate, and have low catalytic activitiestowards CPC to be used in industry. We investigated the catalytic mechanism of CPC acylase by the sequence alignment,homology modeling, and active site analysis to a series of CPC acylases from Pseudomonas where some effective mutationshave been reported for activity enhancement. Two CPC acylases coded by the genes acyII and S12 are studied intensivelyfor the interaction between the amino acid residues in the activity region and the substrate CPC based upon thecomplex structure obtained from the homology modeling and molecular docking. Furthermore, the catalytic parameters ofthe two CPC acylases were measured experimentally in order to corroborate the modeling analysis and propose potentialdesigning strategy for improvement of enzymic activity.