Labade Chaitali P, Jadhav Abhilash R, Ahire Mehul, Zinjarde Smita S, Tamhane Vaijayanti A
Institute of Bioinformatics and Biotechnology (IBB), Savitribai Phule Pune University, (SPPU), Ganeshkhind Road, Pune 411007, Maharashtra, India.
Institute of Bioinformatics and Biotechnology (IBB), Savitribai Phule Pune University, (SPPU), Ganeshkhind Road, Pune 411007, Maharashtra, India.
Ecotoxicol Environ Saf. 2018 Jan;147:612-621. doi: 10.1016/j.ecoenv.2017.09.028. Epub 2017 Oct 10.
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.
本研究探讨了棉铃虫中基于谷胱甘肽 - S - 转移酶(GST)的农药解毒作用及其在从环境中消除农药方面的潜在应用。将农药混合物(有机磷类——毒死蜱和敌敌畏、拟除虫菊酯类——氯氰菊酯;每种浓度为2 - 15ppm)经饮食暴露于棉铃虫幼虫,导致GST活性和基因表达呈剂量依赖性上调。从喂食10ppm农药混合物的幼虫中分离出棉铃虫的一种变异型GST(HaGST - 8),并在酵母(毕赤酵母HaGST - 8)中进行重组表达。HaGST - 8的分子量为29kDa,在30°C、pH 9时活性最高。气相色谱 - 质谱联用(GC - MS)和液相色谱 - 高分辨质谱(LC - HRMS)分析证实,HaGST - 8能有效消除有机磷类农药,并使水溶液中的氯氰菊酯含量部分降低(53%)。与未转化的酵母不同,毕赤酵母HaGST - 8在添加了农药混合物(每种农药200和400ppm)的培养基中生长良好,这表明HaGST - 8具有解毒能力。HaGST - 8的氨基酸序列与已报道的HaGST - 7序列仅有2个错配。分子相互作用强度研究表明,HaGST - 8与有机磷类、拟除虫菊酯类、有机氯类、氨基甲酸酯类和新烟碱类农药具有更强的结合亲和力。重组HaGST - 8消除农药的能力以及毕赤酵母HaGST - 8在高农药含量环境中大量生长的能力可应用于从食物、水资源中去除此类残留物以及生物修复。