Abstract
A novel esterase gene, pytH, encoding a pyrethroid-hydrolyzing carboxylesterase was cloned from Sphingobium sp. strain JZ-1. The gene contained an open reading frame of 840 bp. Sequence identity searches revealed that the deduced enzyme shared the highest similarity with many alpha/beta-hydrolase fold proteins (20 to 24% identities). PytH was expressed in Escherichia coli BL21 and purified using Ni-nitrilotriacetic acid affinity chromatography. It was a monomeric structure with a molecular mass of approximately 31 kDa and a pI of 4.85. PytH was able to transform p-nitrophenyl esters of short-chain fatty acids and a wide range of pyrethroid pesticides, and isomer selectivity was not observed. No cofactors were required for enzyme activity.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Carboxylesterase / chemistry
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Carboxylesterase / genetics*
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Carboxylesterase / isolation & purification
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Carboxylesterase / metabolism*
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Chromatography, Affinity / methods
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Cloning, Molecular
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DNA, Bacterial / chemistry
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DNA, Bacterial / genetics
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Escherichia coli / genetics
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Fatty Acids, Volatile / metabolism
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Isoelectric Point
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Molecular Sequence Data
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Molecular Weight
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Open Reading Frames
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Pyrethrins / metabolism*
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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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Sphingomonadaceae / enzymology*
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Sphingomonadaceae / genetics
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Substrate Specificity
Substances
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DNA, Bacterial
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Fatty Acids, Volatile
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Pyrethrins
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Recombinant Proteins
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Carboxylesterase
Associated data
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GENBANK/FJ686047
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GENBANK/FJ688006