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从二化螟(Chilo suppressalis Walker,鳞翅目:螟蛾科)中肠和唾液腺提取的α-和β-葡萄糖苷酶的酶学特性

Enzymatic properties of alpha- and beta-glocusidases extracted from midgut and salivary glands of rice striped stem borer, Chilo suppressalis Walker (Lepidoptera: Pyralidae).

作者信息

Zibaee Arash, Bandani Ali Reza, Ramzi Samar

机构信息

Department of Plant Protection, College of agriculture and natural resources, University of Tehran, Karaj 31584, Iran.

出版信息

C R Biol. 2009 Jul;332(7):633-41. doi: 10.1016/j.crvi.2009.02.009. Epub 2009 May 9.

Abstract

The study of digestive enzymes, especially in important pests like Chilo suppressalis Walker (Lepidoptera: Pyralidae), which are a key constraint on rice production in a wide area of the globe and also in Iran, could be a successful procedure in the development of a safe and useful control strategy. Glycosidase are a type of digestive enzymes which have a critical role in the final stages of carbohydrate digestion; they hydrolyze alpha-D-(1,4)-glucose linkage such as p-nitrophenyl-alpha-D-glucoside in di and oligosaccharide components. Laboratory reared 4th instar larvae were randomly selected; midgut and salivary gland were removed by dissection under a stereo microscope and glucosidase activities were assayed by Ferreira and Terra's procedures. The activities of alpha- and beta-glucosidase in the midgut and salivary gland were 0.009, 0.0063, 0.005 and 0.003 micromol/min/mg protein, respectively. The optimal pH and temperature for enzyme activity were determined to be 9 and 45 degrees C for the glucosidases measured, values which are in agreement with other reports, especially in lepidopteran insects, which give values between 8-12 and 20-50 degrees C. The enzyme activity increased with the addition of NaCl, MgCl(2) and CaCl(2) and decreased due to the use of different concentrations of KCl, Urea, EDTA, SDS and Urea both in midgut and the salivary glands. Control of pests by using resistant varieties is one of the most important practices that are dependent on inhibitors in plants. Hence, characterization of digestive enzymes, especially the effect of inhibitors on enzyme activity, could be useful, on the one hand for a better understanding of enzyme roles in the nutrition physiology of insects, and on the other hand to reach safe and useful controls of insect pests.

摘要

对消化酶的研究,尤其是在诸如二化螟(鳞翅目:螟蛾科)等重要害虫中的研究,二化螟是全球广大地区以及伊朗水稻生产的关键制约因素,这可能是制定安全且有效的控制策略的一个成功方法。糖苷酶是一类消化酶,在碳水化合物消化的最后阶段起着关键作用;它们水解二糖和寡糖成分中的α-D-(1,4)-葡萄糖键,如对硝基苯基-α-D-葡萄糖苷。随机选取实验室饲养的四龄幼虫;在体视显微镜下解剖取出中肠和唾液腺,并按照费雷拉和特拉的方法测定葡糖苷酶活性。中肠和唾液腺中α-和β-葡糖苷酶的活性分别为0.009、0.0063、0.005和0.003微摩尔/分钟/毫克蛋白质。所测葡糖苷酶的酶活性最佳pH值和温度分别确定为9和45摄氏度,这些值与其他报告一致,特别是在鳞翅目昆虫中,其值在8 - 12摄氏度和20 - 50摄氏度之间。在中肠和唾液腺中,酶活性随着NaCl、MgCl₂和CaCl₂的添加而增加,而由于使用不同浓度的KCl、尿素、EDTA、SDS和尿素而降低。利用抗性品种控制害虫是依赖植物中抑制剂的最重要措施之一。因此,对消化酶进行表征,尤其是抑制剂对酶活性的影响,一方面有助于更好地理解酶在昆虫营养生理学中的作用,另一方面有助于实现对害虫的安全有效控制。

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