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Imidacloprid is degraded by CYP353D1v2, a cytochrome P450 overexpressed in a resistant strain of Laodelphax striatellus.
Pest Manag Sci. 2017 Jul; 73(7):1358-1363.PM

Abstract

BACKGROUND

Cytochrome P450s are associated with the metabolising of a wide range of compounds, including insecticides. CYP353D1v2 has been found to be overexpressed in an imidacloprid-resistant strain of Laodelphax striatellus. Thus, this study was conducted to express CYP353D1v2 in Sf9 cells as a recombinant protein, to assess its ability to metabolise imidacloprid.

RESULTS

Western blot and carbon monoxide difference spectrum analysis indicated that the intact CYP353D1v2 protein had been successfully expressed in Sf9 insect cells. Catalytic activity tests with four traditional P450-activity-probing substrates found that the expressed CYP353D1v2 preferentially metabolised p-nitroanisole, ethoxycoumarin and ethoxyresorufin with specific activities of 32.70, 0.317 and 1.22 pmol min-1 pmol-1 protein respectively, but no activity to luciferin-H EGE. The enzyme activity for degrading imidacloprid was tested by measuring substrate depletion and formation of the metabolite. Kinetic parameters for imidacloprid were Km 5.99 ± 0.95 µm and kcat 0.03 ± 0.0004 min-1 . The chromatogram analysis showed clearly the NADPH-dependent depletion of imidacloprid and the formation of an unknown metabolite. The UPLC-MS mass spectrum demonstrated that the metabolite was an oxidative product of imidacloprid, 5-hydroxy-imidacloprid.

CONCLUSION

These results suggest that CYP353D1v2 in L. striatellus is capable of degrading imidacloprid, and that enzyme activity can be evaluated well only by some traditional probing substrates. © 2017 Society of Chemical Industry.

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Authors+Show Affiliations

Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.

Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.

Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.

Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.

Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.

State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, Jiangsu, China.

Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.

MeSH

AnimalsCytochrome P-450 Enzyme SystemHemipteraInactivation, MetabolicInsect ProteinsInsecticide ResistanceInsecticidesNeonicotinoidsNitro CompoundsSf9 CellsSpodoptera

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28296046

Citation

Elzaki, Mohammed Esmail Abdalla, et al. "Imidacloprid Is Degraded By CYP353D1v2, a Cytochrome P450 Overexpressed in a Resistant Strain of Laodelphax Striatellus." Pest Management Science, vol. 73, no. 7, 2017, pp. 1358-1363.

Elzaki MEA, Miah MA, Wu M, et al. Imidacloprid is degraded by CYP353D1v2, a cytochrome P450 overexpressed in a resistant strain of Laodelphax striatellus. Pest Manag Sci. 2017;73(7):1358-1363.

Elzaki, M. E. A., Miah, M. A., Wu, M., Zhang, H., Pu, J., Jiang, L., & Han, Z. (2017). Imidacloprid is degraded by CYP353D1v2, a cytochrome P450 overexpressed in a resistant strain of Laodelphax striatellus. Pest Management Science, 73(7), 1358-1363. https://doi.org/10.1002/ps.4570

Elzaki MEA, et al. Imidacloprid Is Degraded By CYP353D1v2, a Cytochrome P450 Overexpressed in a Resistant Strain of Laodelphax Striatellus. Pest Manag Sci. 2017;73(7):1358-1363. PubMed PMID: 28296046.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Imidacloprid is degraded by CYP353D1v2, a cytochrome P450 overexpressed in a resistant strain of Laodelphax striatellus. AU - Elzaki,Mohammed Esmail Abdalla, AU - Miah,Mohammad Asaduzzaman, AU - Wu,Min, AU - Zhang,Haomiao, AU - Pu,Jian, AU - Jiang,Ling, AU - Han,Zhaojun, Y1 - 2017/04/24/ PY - 2017/01/05/received PY - 2017/02/13/revised PY - 2017/03/06/accepted PY - 2017/3/16/pubmed PY - 2018/7/7/medline PY - 2017/3/16/entrez KW - cytochrome P450 KW - functional expression KW - imidacloprid detoxification KW - insecticide metabolism KW - probing substrates SP - 1358 EP - 1363 JF - Pest management science JO - Pest Manag Sci VL - 73 IS - 7 N2 - BACKGROUND: Cytochrome P450s are associated with the metabolising of a wide range of compounds, including insecticides. CYP353D1v2 has been found to be overexpressed in an imidacloprid-resistant strain of Laodelphax striatellus. Thus, this study was conducted to express CYP353D1v2 in Sf9 cells as a recombinant protein, to assess its ability to metabolise imidacloprid. RESULTS: Western blot and carbon monoxide difference spectrum analysis indicated that the intact CYP353D1v2 protein had been successfully expressed in Sf9 insect cells. Catalytic activity tests with four traditional P450-activity-probing substrates found that the expressed CYP353D1v2 preferentially metabolised p-nitroanisole, ethoxycoumarin and ethoxyresorufin with specific activities of 32.70, 0.317 and 1.22 pmol min-1 pmol-1 protein respectively, but no activity to luciferin-H EGE. The enzyme activity for degrading imidacloprid was tested by measuring substrate depletion and formation of the metabolite. Kinetic parameters for imidacloprid were Km 5.99 ± 0.95 µm and kcat 0.03 ± 0.0004 min-1 . The chromatogram analysis showed clearly the NADPH-dependent depletion of imidacloprid and the formation of an unknown metabolite. The UPLC-MS mass spectrum demonstrated that the metabolite was an oxidative product of imidacloprid, 5-hydroxy-imidacloprid. CONCLUSION: These results suggest that CYP353D1v2 in L. striatellus is capable of degrading imidacloprid, and that enzyme activity can be evaluated well only by some traditional probing substrates. © 2017 Society of Chemical Industry. SN - 1526-4998 UR - https://www.unboundmedicine.com/medline/citation/28296046/Imidacloprid_is_degraded_by_CYP353D1v2_a_cytochrome_P450_overexpressed_in_a_resistant_strain_of_Laodelphax_striatellus_ L2 - https://doi.org/10.1002/ps.4570 DB - PRIME DP - Unbound Medicine ER -