Enzyme-substrate interaction and characterization of a 2,3- dihydroxybiphenyl 1,2-dioxygenase from Dyella ginsengisoli LA-4

Abstract

A bphC gene (915 bp) encoding 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC) was amplified by PCR from Dyella ginsengisoli LA-4, which was heterologously expressed in Escherichia coli. The purified His-Tag BphC was able to catalyze the meta-cleavage reaction of the dihydroxylated aromatic rings. According to the specificity constant (Kcat/Km) of BphC-LA-4, the specificity of BphC-LA-4 was determined in the following order: 2,3-dihydroxybiphenyl>3-methylcatechol>catechol>4-chlorocatechol>4- methylcatechol. The experimental data were consistent with the prediction of enzyme-substrate complexes. The highest specific activity of BphC-LA-4 was 118.3 U mg-1 for 2,3-dihydroxybiphenyl. © 2009 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.