Role of induced glutathione-S-transferase from Helicoverpa armigera (Lepidoptera: Noctuidae) HaGST-8 in detoxification of pesticides
Abstract The present study deals with glutathione-S-transferase (GST) based detoxification of pesticides in Helicoverpa armigera and its potential application in eliminating pesticides from the environment. Dietary exposure of a pesticide mixture (organophosphates – chlorpyrifos and dichlorvos, pyrethroid – cypermethrin; 2–15ppm each) to H. armigera larvae resulted in a dose dependant up-regulation of GST activity and gene expression. A variant GST from H. armigera ( HaGST-8 ) was isolated from larvae fed with 10ppm pesticide mixture and it was recombinantly expressed in yeast ( Pichia pastoris HaGST-8). HaGST-8 had a molecular mass of 29kDa and was most active at pH 9 at 30°C. GC-MS and LC-HRMS analysis validated that HaGST-8 was effective in eliminating organophosphate type of pesticides and partially reduced the cypermethrin content (53%) from aqueous solutions. Unlike the untransformed yeast, P. pastoris HaGST-8 grew efficiently in media supplemented with pesticide mixtures (200 and 400ppm each pesticide) signifying the detoxification ability of HaGST-8. The amino acid sequence of HaGST-8 and the already reported sequence of HaGST-7 had just 2 mismatches. The studies on molecular interaction strengths revealed that HaGST-8 had stronger binding affinities with organophosphate, pyrethroid, organochloride, carbamate and neonicotinoid type of pesticides. The abilities of recombinant HaGST-8 to eliminate pesticides and P. pastoris HaGST-8 to grow profusely in the presence of high level of pesticide content can be applied for removal of such residues from food, water resources and bioremediation. Highlights Dietary exposure to pesticide mix led to a dose dependant up-regulation of GST activity in pest Helicoverpa armigera . Recombinant protein of HaGST-8 cloned from H. armigera larvae exposed to 10 ppm pesticide mixture was active at alkaline pH. HaGST-8 is very efficient in reducing organophosphate and pyrethroid pesticides. HaGST-8 confers high level of pesticide tolerance capacity in P.pastoris . In silico studies, interaction strength between HaGST-8/HaGST-7 and pesticides indicated variable binding affinities. Graphical abstract [DISPLAY OMISSION]
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