Effect of a functional recombinant cytochrome P450 enzyme of Helicoverpa armigera on gossypol metabolism co-expressed with NADPH-cytochrome P450 reductase in Pichia pastoris
文献类型： 外文期刊
作者： Chen, Cheng 1 ; Pi, Wenhui 2 ; Zhang, Yan 3 ; Nie, Cun-xi 1 ; Liang, Jing 1 ; Ma, Xi 1 ; Wang, Yongqiang 1 ; Ge, Wenxia 1 ;
作者机构： 1.Shihezi Univ, Coll Anim Sci & Technol, Shihezi 832000, Xinjiang, Peoples R China
2.Xinjiang Acad Agr & Reclamat Sci, State Key Lab Sheep Genet Improvement & Hlth Prod, Shihezi 832000, Xinjiang, Peoples R China
3.Shihezi Univ, Sch Chem & Chem Engn, Shihezi 832000, Xinjiang, Peoples R China
4.China Agr Univ, Coll Anim Sci & Technol, State Key Lab Anim Nutr, Beijing 100193, Peoples R China
关键词： Helicoverpa armigera; Cytochrome 9450; CYP6AE14; CYP9A12; Gossypol; Detoxification
期刊名称：PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY （ 影响因子：3.963； 五年影响因子：4.454 ）
ISSN： 0048-3575
年卷期： 2019 年 155 卷
页码：
收录情况： SCI
摘要： Gossypol is a polyphonic toxic compound that is present in cotton plants. The P450 cytochromes CYP6AE14 and CYP9A12 of Helicoverpa armigera are highly induced by gossypol and have been reported to be possibly involved in gossypol degradation. To determine whether the candidate H. armigera CYP6AE14 and CYP9A12 enzymes could metabolize gossypol in vitro, functional recombinant H. armigera CYP6AE14 and CPR (CYP9A12 and CPR) enzymes were successfully expressed in Pichia pastoris (P. pastoris). UPLC-QTOF/MS demonstrated the following results: (1) Free gossypol was spontaneously degraded to the gossypol metabolites G1 (m/z 265) and G2 (m/z 293) without the addition of any enzyme. (2) Free gossypol was observed following the addition of the endogenous or recombinant H. armigera P450 cytochrome CYP6AE14/CYP9A12 enzyme: in the first pathway, free gossypol was dehydroxylated and decarboxylated to G3 (m/z 453), and in the second pathway, the aldehyde group of gossypol and its metabolite were covalently bound with the amine products to form G4 (m/z 437) and G5 (m/z 783). (3) In addition to the gossypol binding pathways, the recombinant H. armigera CPR and CYP9Al2 enzymes was found that could further decarboxylate the gossypol intermediate demethylated reduction of gossypolonic acid (m/z 294) and demethylated gossic acid (m/z 265) to GO (m/z 209) and GO' (m/z 249) respectively.
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