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Digestive proteinase activity in corn earworm (helicoverpa zea) after molting and in response to lowered redox potential

机译:玉米穗虫(helicoverpa zea)蜕皮后和对氧化还原电位降低的反应中的消化蛋白酶活性

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Insect digestive proteinases are often strongly influenced by ambient physicochemical conditions, such as pH, ionic strength, and oxidation-reduction potential. Although the effects of the former two parameters are well documented, the influence ofredox potential on catalytic rates of digestive enzymes is not well understood. In this study, we manipulated the midgut redox potential of a generalist caterpillar (the corn earworm, Helicoverpa zea) by augmenting artificial diet with dithio-threitol,a powerful thiol reducing agent that lowers the redox potential in the lumen by 40-45 mV. Effects on total proteolytic activity, as well as on elastase, chymotrypsin, trypsin, leucine aminopeptidase, and carboxypeptidase A and B activities were measured using azocasein and nitroanilide model substrates. The profiles of proteinase activities in the epithelium and lumen were also monitored on days 1, 2, and 3 after the molt in penultimate instar larvae. Although the reducing agent strongly inhibited the activity of some proteinases in vitro, ingestion of the reducing diet failed to affect in vivo proteinase activities. There was also on effect on larval relative growth, consumption, or digestive efficiencies. We conclude that dietary reducing agentsmust lower midgut redox potential to below -40 mV to significantly impact digestive efficiency.
机译:昆虫的消化蛋白酶通常受周围物理化学条件的强烈影响,例如pH,离子强度和氧化还原电位。尽管前两个参数的影响已得到很好的证明,但还没有很好地了解氧化还原电势对消化酶催化速率的影响。在这项研究中,我们通过用二硫苏糖醇增加人工饮食来控制通才毛毛虫(玉米耳虫,Helicoverpa zea)的中肠氧化还原电位,二硫苏糖醇是一种强大的硫醇还原剂,可将内腔中的氧化还原电位降低40-45 mV。使用偶氮酪蛋白和硝基苯胺模型底物测量对总蛋白水解活性以及对弹性蛋白酶,胰凝乳蛋白酶,胰蛋白酶,亮氨酸氨基肽酶以及羧肽酶A和B活性的影响。在倒数第二龄幼虫蜕皮后的第1、2和3天,还监测了上皮和管腔中蛋白酶活性的概况。尽管还原剂在体外强烈地抑制了某些蛋白酶的活性,但是摄取还原饮食不能影响体内蛋白酶的活性。对幼虫的相对生长,消耗或消化效率也有影响。我们得出结论,饮食还原剂必须将中肠氧化还原电位降低至-40 mV以下,以显着影响消化效率。

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