Effect of a functional recombinant cytochrome P450 enzyme of Helicoverpa armigera on gossypol metabolism co-expressed with NADPH-cytochrome P450 reductase in Pichia pastoris
| Autor: | Xi Ma, Wenju Zhang, Cheng Chen, Wenxia Ge, Yongqiang Wang, Wenhui Pi, Cunxi Nie, Jing Liang, Yan Zhang |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine Health Toxicology and Mutagenesis Metabolite Moths Helicoverpa armigera 01 natural sciences Pichia law.invention Pichia pastoris 03 medical and health sciences chemistry.chemical_compound Cytochrome P-450 Enzyme System law Animals NADPH-Ferrihemoprotein Reductase chemistry.chemical_classification biology fungi Gossypol Cytochrome P450 General Medicine Metabolism biology.organism_classification 010602 entomology 030104 developmental biology Enzyme Biochemistry chemistry Recombinant DNA biology.protein Agronomy and Crop Science |
| Zdroj: | Pesticide Biochemistry and Physiology. 155:15-25 |
| ISSN: | 0048-3575 |
| DOI: | 10.1016/j.pestbp.2019.01.003 |
| Popis: | 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 CYP9A12 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 G0 (m/z 209) and G0' (m/z 249) respectively. |
| Databáze: | OpenAIRE |
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