High-Level Expression and Secretion of Methyl Parathion Hydrolase in Bacillus subtilis WB800

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High-level expression and secretion of methyl parathion hydrolase in Bacillus subtilis WB800.

The methyl parathion hydrolase (MPH)-encoding gene mpd was placed under the control of the P43 promoter and Bacillus subtilis nprB signal peptide-encoding sequence. High-level expression and secretion of mature, authentic, and stable MPH were achieved using the protease-deficient strain B. subtilis WB800 as the host.

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Isolation of methyl parathion-degrading strain M6 and cloning of the methyl parathion hydrolase gene.

A degradative bacterium, M6, was isolated and presumptively identified as Plesiomonas sp. strain M6 was able to hydrolyze methyl parathion to p-nitrophenol. A novel organophosphate hydrolase gene designated mpd was selected from its genomic library prepared by shotgun cloning. The nucleotide sequence of the mpd gene was determined. The gene could be effectively expressed in Escherichia coli.

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BACKGROUND Although Pichia pastoris has been successfully used to produce various recombinant heterologous proteins, the efficiency varies. In this study, we used methyl parathion hydrolase (MPH) from Ochrobactrum sp. M231 as an example to study the effect of protein amino acid sequence on secretion from P. pastoris. RESULTS The results indicated that the protein N-terminal sequence, the endo...

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ژورنال

عنوان ژورنال: Applied and Environmental Microbiology

سال: 2005

ISSN: 0099-2240,1098-5336

DOI: 10.1128/aem.71.7.4101-4103.2005