Expression of organophosphohydrolase in recombinant Escherichia coli and its purification and immobilization by one-step method

ZHOU Li-Ya1 OUYANG Ya-Ping1 XUE Sai-Guang1 JIANG Yan-Jun1

(1.School of Chemical Engineering and Technology, Hebei University of Technology, Tianjin, China 300130)

【Abstract】[Background] As a kind of broad-spectrum insecticides, organophosphorus compounds have caused serious environmental pollution due to their high dosage, strong toxicity and non-degradability. [Objective] To degrade the organophosphorus compounds, organophosphohydrolase (OpdA) was immobilized on NiCo2O4 and the properties of the catalyst in degrading organophosphorus compounds were investigated. [Methods] Specifically, histidine tag (His-tag) was introduced into OpdA to construct His-tagged OpdA which was then efficiently expressed in Escherichia coli Rosetta(DE3) using pET-28a(+) as vector under the induction of 1.0 mmol/L isopropyl-β-D-thiogalactopyranoside. The immobilized OpdA (OpdA@NiCo2O4) was prepared by one-step purification and immobilization method. [Results] NiCo2O4 was prepared by hydrothermal treatment and calcination. Based on the coordination between the transition metal ions on the surface of NiCo2O4 and the imidazole group of histidine on the surface of enzyme, one-step purification and immobilization of His-tagged OpdA were achieved. OpdA@NiCo2O4 with high stability was obtained under optimized conditions, and then it was applied for the degradation of organophosphorus compounds. The high degradation efficiency of organophosphorus compounds was realized by cascade reaction in the presence of NaBH4 under the optimal degradation conditions. [Conclusion] In conclusion, one-step purification and immobilization of recombinant enzyme were achieved, which proved that this study provided a safe, efficient and environmentally friendly new way for the degradation of organophosphorus compounds.

【Keywords】 Organophosphohydrolase; Recombinant expression; His-tag; One-step purification and immobilization; Organophosphorus compound;


【Funds】 National Natural Science Foundation of China (21878068) Natural Science Foundation of Hebei Province (B2017202056) Program for Top 100 Innovative Talents in Colleges and Universities of Hebei Province (SLRC2017029) Hebei High Level Personnel of Support Program (A2016002027)

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This Article


CN: 11-1996/Q

Vol 47, No. 07, Pages 2094-2105

July 2020


Article Outline


  • 1 Materials and methods
  • 2 Results
  • 3 Discussion and conclusion
  • References