Abstract
A 2,3-dihydroxybiphenyl-1,2-dioxygenase gene (designated as bphC_meta) was identified in activated sludge metagenome by PCR. This gene shared 99% sequence identity with BphC from Burkholderia xenovorans LB400. The enzyme was purified from recombinant Escherichia coli with a subunit molecular mass of 32 ± 1 kDa. It was optimally active at pH 9.0 and 40°C, using 2,3-dihydroxybiphenyl as a substrate. Activity toward substituted catechols was: 2,3-dihydroxybiphenyl > 3-methylcatechol > catechol > 4-chlorocatechol (4-methylcatechol). The prediction made by molecular docking was consistent with the kinetic experimental data, and further explained the substrate preference of BphC_meta. The present study could pave the way for the improved understanding and application of BphCs derived from metagenomes.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Biphenyl Compounds / metabolism*
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Burkholderia / enzymology
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Burkholderia / genetics
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Catechols / metabolism*
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Dioxygenases / genetics*
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Dioxygenases / isolation & purification
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Dioxygenases / metabolism*
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Enzyme Stability
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Escherichia coli / genetics
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Hydrogen-Ion Concentration
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Kinetics
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Metagenome*
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Molecular Dynamics Simulation
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Molecular Sequence Data
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Molecular Weight
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Polymerase Chain Reaction
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Sewage / microbiology*
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Substrate Specificity
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Temperature
Substances
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Biphenyl Compounds
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Catechols
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Recombinant Proteins
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Sewage
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2,3-dihydroxybiphenyl
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Dioxygenases
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2,3-dihydroxybiphenyl oxygenase