title: Comparative Analysis of Recombinant Cytochrome P450 CYP9A61 from Cydia pomonella Expressed in Escherichia coli and Pichia pastoris.
abstract: On the basis of prior work, cytochrome P450 CYP9A61 was found to be enriched in fat bodies and during feeding stages, and transcription was induced by λ-cyhalothrin in Cydia pomonella. In this study, recombinant CYP9A61 was expressed in Escherichia coli and Pichia pastoris, and its biochemical properties were investigated. Substrate saturation curves and biochemical properties revealed that, in the presence of glycosylation, the yeast-secreted CYP9A61 exhibited a higher affinity for the substrate p-nitroanisole and was found to be more stable at certain pHs and temperatures than bacterially produced CYP9A61. Half-inhibitory concentrations (IC50) of three synthetic pyrethroids on both the bacterium- and yeast-expressed CYP9A61 suggested that recombinant CYP9A61 expressed in different hosts exhibits different inhibition properties. Taken together, our findings show that yeast-expressed CYP9A61 exhibits enzyme activity that is better than that expressed in bacteria and might be used for further metabolism assays to reveal the insecticide-detoxifying role of CYP9A61 in C. pomonella.
authors.affiliation: Key Laboratory of Economical and Applied Entomology of Liaoning Province, College of Plant Protection, Shenyang Agricultural University , Shenyang 110866, China.
authors.forename: Xue-Qing
authors.identifier: http://orcid.org/0000-0002-3919-8013
authors.initials: XQ
authors.lastname: Yang
authors.affiliation: Key Laboratory of Economical and Applied Entomology of Liaoning Province, College of Plant Protection, Shenyang Agricultural University , Shenyang 110866, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Institute of Plant Protection, Chinese Academy of Agricultural Sciences , Beijing 100193, China.
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Tan
authors.affiliation: Key Laboratory of Economical and Applied Entomology of Liaoning Province, College of Plant Protection, Shenyang Agricultural University , Shenyang 110866, China.
authors.forename: Xiao-Qi
authors.initials: XQ
authors.lastname: Wang
authors.affiliation: Key Laboratory of Economical and Applied Entomology of Liaoning Province, College of Plant Protection, Shenyang Agricultural University , Shenyang 110866, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Dong
mesh_terms: D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D007915:Lepidoptera / Q000201:enzymology* / Q000235:genetics; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature
chemical_list: D019476:Insect Proteins; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System
keywords: Cydia pomonella (L.); P450; biochemical properties; detoxification; inhibition properties; pyrethroids