Abstract
CYP353D1v2 is a cytochrome P450 related to imidacloprid resistance in Laodelphax striatellus. This work was conducted to examine the ability of CYP353D1v2 to metabolize other insecticides. Carbon monoxide difference spectra analysis indicates that CYP353D1v2 was successfully expressed in insect cell Sf9. The catalytic activity of CYP353D1v2 relating to degrading buprofezin, chlorpyrifos, and deltamethrin was tested by measuring substrate depletion and analyzing the formation of metabolites. The results showed the nicotinamide–adenine dinucleotide phosphate (NADPH)-dependent depletion of buprofezin (eluting at 8.7 min) and parallel formation of an unknown metabolite (eluting 9.5 min). However, CYP353D1v2 is unable to metabolize deltamethrin and chlorpyrifos. The recombinant CYP353D1v2 protein efficiently catalyzed the model substrate p-nitroanisole with a maximum velocity of 9.24 nmol/min/mg of protein and a Michaelis constant of Km = 6.21 µM. In addition, imidacloprid was metabolized in vitro by the recombinant CYP353D1v2 microsomes (catalytic constant Kcat) 0.064 pmol/min/pmol P450, Km = 6.41 µM. The mass spectrum of UPLC-MS analysis shows that the metabolite was a product of buprofezin, which was buprofezin sulfone. This result provided direct evidence that L. striatellus cytochrome P450 CYP353D1v2 is capable of metabolizing imidacloprid and buprofezin.
Highlights
Buprofezin is an insect growth regulator insecticide, mainly used to control homopteran insect pests
Their function is related to the metabolism of a broad range of xenobiotics, including insecticides [9]
Little is known about its substrate specificity, and whether the crosswhether the cross-resistance among insecticides of different by a in single or resistance among insecticides of different kinds is mediated by akinds singleisormediated multiple P450s insects
Summary
Buprofezin is an insect growth regulator insecticide, mainly used to control homopteran insect pests. A great deal of research has been conducted to study the molecular usually developed by two major mechanisms; metabolic resistance by enhanced detoxification mechanism of insecticide resistance in L. striatellus. Buprofezin resistance has been attributed to major mechanisms; metabolic resistance by enhanced detoxification enzymes and target-site insensitive metabolic resistance due to the increase of the expression level of a cytochrome P450 gene in mutations. Their function is related to the metabolism of a broad range of xenobiotics, including insecticides [9]. A superfamily of genes in different organisms Their function is related to the metabolism of a broad. Few insecticide resistance-related genes have been studied for their substrate spectrum.
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