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Cloning, expression, purification, and characterization of an organophosphate-degrading enzyme in Escherichia coli

文献类型: 会议论文

第一作者: Jinbin Wang

作者: Jinbin Wang 1 ; Dachao Chen 1 ; Dongxiao Hu 1 ; Xuefeng Su 1 ; Xueming Tang 1 ;

作者机构: 1.Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences

关键词: Organophosphate-degrading enzyme;Expression;Purification;Characterization

会议名称: International Conference on Energy, Environment and Sustainable Development

主办单位:

页码: 201-207

摘要: OpdA is one of organic phosphorus degradation enzyme gene from Agrobacterium radiobacter that may be the most promising targets for the digestion of digestion. In this study, we describe for the cloning and expression in Escherichia coli (E. coli) of plasmid pET28b-opdA, followed by purification by NTA-Ni~(2+) agarose affinity chromatography. Protein expression and purification were evaluated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The Results showed that the optimal value of inoculum OD600 before induction, inducing time, final IPTG concentration and inducing temperature respectively were 0.5, 5 h, 1 mmol/L, 37℃. We obtained the concentration of renatured protein was 18.312 mg/L. The Km was 4.26μmol/L at 37℃, and the maximum reaction velocity (Vmax) was 3.2669μmol/L·min.

分类号: x1-532

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