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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins / genetics*
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism*
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Cloning, Molecular
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DNA, Bacterial / chemistry
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DNA, Bacterial / genetics
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Dioxygenases / genetics*
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Dioxygenases / isolation & purification
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Dioxygenases / metabolism*
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Escherichia coli / genetics
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Kinetics
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Models, Molecular
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Molecular Sequence Data
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Phylogeny
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Protein Structure, Tertiary
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Sequence Alignment
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Substrate Specificity
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Xanthomonadaceae / enzymology*
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Xanthomonadaceae / genetics
Substances
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Bacterial Proteins
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DNA, Bacterial
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Dioxygenases
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2,3-dihydroxybiphenyl oxygenase