Department of Biochemistry, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad 431 004, M.S., India.
Saudi J Biol Sci. 2012 Jul;19(3):317-23. doi: 10.1016/j.sjbs.2012.03.001. Epub 2012 Mar 19.
Exploring bacterial communities with proteolytic activity from the gut of the Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) insect pests was the purpose of this study. As initial efforts to achieve this goal here we report the isolation of new Bacillus subtillis RTSBA6 6.00 strain from the gut of H. armigera and demonstrated as proteases producer. Zymographic analysis revealed 12 proteolytic bands with apparent molecular weights ranging from 20 to 185 kDa. Although some activity was detected at acidic pH, the major activity was observed at slight alkaline pH (7.8). The optimum temperature was found to be 35 °C with complete loss of activity at 70 °C. All proteases were completely inactivated by PMSF (phenylmethylsulfonyl fluoride) and TPCK (N-tosyl-l-phenylalanine chloromethyl ketone), suggesting that proteases secreted by B. subtillis RTSBA6 6.00 belong to serine proteases class with chymotrypsin-like activity. The occurrence of protease producing bacterial community in the gut of the H. armigera advocates its probable assistance to insect in proteinaceous food digestion and adaptation to protease inhibitors of host plants.
本研究旨在探索鳞翅目夜蛾科(Lepidoptera:Noctuidae)害虫棉铃虫(Helicoverpa armigera)(Hubner)肠道中具有蛋白水解活性的细菌群落。作为实现这一目标的初步努力,我们在这里报道了从棉铃虫肠道中分离到的新型枯草芽孢杆菌 RTSBA6 6.00 菌株,并证明其具有蛋白酶产生能力。酶谱分析显示有 12 条蛋白水解带,其表观分子量范围为 20 至 185 kDa。尽管在酸性 pH 值下检测到一些活性,但主要活性是在轻微碱性 pH 值(7.8)下观察到的。最适温度为 35°C,70°C 时活性完全丧失。所有蛋白酶均被 PMSF(苯甲基磺酰氟)和 TPCK(N-甲苯磺酰-L-苯丙氨酸氯甲基酮)完全失活,表明枯草芽孢杆菌 RTSBA6 6.00 分泌的蛋白酶属于丝氨酸蛋白酶类,具有胰凝乳蛋白酶样活性。蛋白酶产生细菌群落存在于棉铃虫肠道中,这表明它可能有助于昆虫对蛋白质食物的消化,并适应宿主植物的蛋白酶抑制剂。