Abstract
In silico substrate docking of both stereoisomers of the pesticide chlorfenvinphos (CVP) in the phosphotriesterase from Agrobacterium radiobacter identified two residues (F131 and W132) that prevent productive substrate binding and cause stereospecificity. A variant (W131H/F132A) was designed that exhibited ca. 480-fold and 8-fold increases in the rate of Z-CVP and E-CVP hydrolysis, respectively, eliminating stereospecificity.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Substitution / genetics*
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Catalytic Domain / genetics*
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Chlorfenvinphos / metabolism*
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Models, Molecular
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Mutagenesis, Site-Directed
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Phosphoric Triester Hydrolases / chemistry
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Phosphoric Triester Hydrolases / genetics*
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Phosphoric Triester Hydrolases / metabolism*
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Protein Structure, Tertiary
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Rhizobium / enzymology*
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Rhizobium / genetics*
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Substrate Specificity
Substances
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Recombinant Proteins
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Phosphoric Triester Hydrolases
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Chlorfenvinphos