期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 57, 期 19, 页码 8901-8905出版社
AMER CHEMICAL SOC
DOI: 10.1021/jf901739g
关键词
Tat pathway; twin-arginine signal peptide; periplasmic secretion; methyl parathion hydrolase
资金
- 863 Hi-Tech Research and Development [2007AA06Z335, 2007AA061101]
The uptake of organophosphates (OPs) is a rate-limiting factor in whole-cell biocatalysis systems. Here, we report the periplasmic secretion of methyl parathion hydrolase (MPH) by employing the twin-arginine translocation (Tat) pathway in Escherichia coli. The twin-arginine signal peptide of trimethylamine N-oxide reductase (TorA) from E coli was used for exporting MPH to the periplasm of E coli, alleviating the substrate uptake limitation. A periplasmic expression vector, pUTM18, coding for TorA-MPH was constructed, and the periplasmic secretion and functionality of MPH were demonstrated by cell fractionation, immunoblotting, and enzyme activity assays. The strain expressing periplasmic MPH showed 3-fold higher whole-cell activity than the control strain expressing cytoplasmic MPH. Suspended cultures also exhibited good stability, retaining almost 100% activity over a period of 2 weeks. Owing to their high activity and superior stability, these live biocatalysts are ideal for large-scale detoxification of OPs.
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