Enzyme–substrate interaction and characterization of a 2,3-dihydroxybiphenyl 1,2-dioxygenase from Dyella ginsengisoli LA-4 |
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| Authors: | Ang Li Yuanyuan Qu Jiti Zhou & Fang Ma |
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| Institution: | Key Laboratory of Industrial Ecology and Environmental Engineering, School of Environmental and Biological Science and Technology, MOE, Dalian University of Technology, Dalian, China;and;School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin, China |
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| 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 ( 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. |
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| Keywords: | Dyella ginsengisoli 2 3-dihydroxybiphenyl 1 2-dioxygenase kinetic parameters enzyme–substrate complex |
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