Abstract
The coding region of mpd gene corresponding to mature methyl parathion hydrolase (MPH) was heterologously overexpressed in Escherichia coli BL21 (DE3) by using pET expression system. The lactose-induced expression yield of MPH is increased 2-fold compared with IPTG as inducer. Furthermore, it was found that specific activity of MPH increased 48% by reducing the induction temperature to 22°C. The addition of 25 mM lactose at 22°C, the MPH activity of fermentation broth had a specific activity of 1.4 × 104 U/mg protein. Plasmid was no significant decrease in the modified medium. The optimal pH and temperature of MPH were 8.0 and 30°C, respectively. Over a period of 5 months, the dried cells showed no significant decrease in the activity of the detoxifying enzymes. The crude enzymes in 50 mM citrate-phosphate buffer (pH 8.0) were able to degrade about 98% of the organophosphate pesticides sprayed on cabbage. The detoxification efficiency was superior to that of the treatments of water, detergent, and a commercially available enzyme product. Additionally, the products of pesticide hydrolysis generated by treatment with the enzyme extract were determined to be virtually nontoxic.
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Acknowledgements
This work was supported by grants from the 863 Hi-Tech Research and Development Program of the People’s Republic of China (No. 2007AA06Z335) and the Innovation Program of the Chinese Academy of Sciences (No. KSCX2-YW-G-008). We appreciate valuable comments and suggestions of reviewers.
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Yang, J., Yang, C., Jiang, H. et al. Overexpression of methyl parathion hydrolase and its application in detoxification of organophosphates. Biodegradation 19, 831–839 (2008). https://doi.org/10.1007/s10532-008-9186-2
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DOI: https://doi.org/10.1007/s10532-008-9186-2