Open AccessEnzyme–substrate interaction and characterization of a 2,3‐dihydroxybiphenyl 1,2‐dioxygenase from Dyella ginsengisoli LA‐4
Author(s) -
Li Ang,
Qu Yuanyuan,
Zhou Jiti,
Ma Fang
Publication year - 2009
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2009.01487.x
Subject(s) - dioxygenase , catechol , enzyme , escherichia coli , chemistry , substrate (aquarium) , stereochemistry , biochemistry , biology , gene , ecology
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 ( K cat / K m ) 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.