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Heterologous expression of Helicoverpa armigera cytochrome P450 CYP6B7 in Pichia pastoris and interactions of CYP6B7 with insecticides
Author(s) -
Zhao Chunqing,
Song Genmiao,
Duan Hongxia,
Tang Tao,
Wang Chen,
Qiu Lihong
Publication year - 2017
Publication title -
pest management science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.296
H-Index - 125
eISSN - 1526-4998
pISSN - 1526-498X
DOI - 10.1002/ps.4552
Subject(s) - helicoverpa armigera , pichia pastoris , fipronil , bifenthrin , heterologous expression , pyrethroid , cytochrome p450 , carboxylesterase , biology , fenvalerate , biochemistry , gene , enzyme , recombinant dna , botany , pesticide , larva , agronomy
BACKGROUND Previous studies indicated that constitutive over‐expression of cytochrome P450 CYP6B7 was involved in fenvalerate resistance in Helicoverpa armigera . In this study, the CYP6B7 gene from H. armigera (namely HaCYP6B7) , was heterologously expressed in Pichia pastoris GS115. A vector pPICZA‐ HaCYP6B7 was constructed and transformed into P. pastoris GS115, the transformant of pPICZA‐ HaCYP6B7 ‐GS115 was then cultured and induced by 1% (v/v) methanol and the heterologous expression of Ha CYP6B7 protein in P. pastoris was confirmed by SDS‐PAGE and western blot. RESULTS Microsomes containing the expressed Ha CYP6B7 showed activities against model substrate p ‐nitroanisole and 7‐ethoxycoumarin, with p ‐nitroanisole O ‐demethylation (PNOD) and 7‐ethoxycoumarin O ‐deethylation (ECOD) activities of 15.66‐ and 4.75‐fold of the control, respectively. Moreover, it showed degradation activities against the insecticides bifenthrin, fenvalerate and chlorpyrifos, with clearance activities of 6.88‐, 1.49‐ and 2.27‐fold of the control, respectively. The interactions of Ha CYP6B7 with insecticides were further confirmed by molecular docking in silico with binding scores of 5.450, 5.295 and 2.197 between putative Ha CYP6B7 protein and bifenthrin, fenvalerate and chlorpyrifos, respectively. CONCLUSION The results of present study provided more direct and important evidence on the role of Ha CYP6B7 conferring pyrethroid resistance in H. armigera . © 2017 Society of Chemical Industry