Cloning and Characterization of mpd and ophc2 from an Organophosphorus Pesticide-degrading Bacteria YC-YH1

doi:10.13560/j.cnki.biotech.bull.1985.2015.11.030

Abstract

Abstract: We cloned two organophosphorus hydrolase genes mpd and ophc2 from an organophosphorus pesticide-degrading bacteria Pseudomonas stutzeri YC-YH1. The two genes were inserted into the expression vector pET-32a and transformed into E. coli BL21(DE3)respectively. We used Ni-affinity column to purify enzymes and then study the nature of the two enzymes. It was suggested that the optimal temperature of MPH is 40℃, and the MPH displays high activity in the pH range of 8 to 12. The optimal temperature of OPHC2 is 30℃. In the pH ranging of 8 to12, OPHC2 has higher activity. The mixture of MPH and OPHC2 in theproportion of1:1 showsed higher activity in the range of 30℃ to 40℃, the enzyme showsed more than 95% activity in the range of pH8-12. A variety of metalions affect the activity of the enzymes and the enzymes are sensitive to SDS and EDTA, but the compound could reduce the sensitivity to the metalions.

Key words: organophosphorus hydrolase gene, MPH, OPHC2, purification, enzymatic property

Jia Yang, Shi Yanhua, Ren Lei, Qiao Cheng, Wang Junhuan, Yan Yanchun. Cloning and Characterization of mpd and ophc2 from an Organophosphorus Pesticide-degrading Bacteria YC-YH1[J]. Biotechnology Bulletin, 2015, 31(11): 228-235.

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