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赤拟谷盗(Panzer)(鞘翅目:拟步甲科)幼虫中肠α-淀粉酶的生化特性

Biochemical Properties of α-Amylase from Midgut of Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) Larvae.

作者信息

Cruz W O, Sinhori G G C, de Lima C A R, Pontes E G

机构信息

Lab de Bioquímica e Biologia Molecular de Artrópodes Depto de Química, Univ Federal Rural do Rio de Janeiro, Br. 465, Km 7, Seropédica, Rio de Janeiro, 23890-000, Brasil.

Instituto de Zootecnia, Univ Federal Rural do Rio de Janeiro, Seropédica, Brazil.

出版信息

Neotrop Entomol. 2018 Oct;47(5):698-708. doi: 10.1007/s13744-018-0590-y. Epub 2018 Feb 26.

Abstract

The lesser mealworm, Alphitobius diaperinus (Panzer), is the main insect pest in the poultry industry, thus causing serious damage to production. In this work, the properties of midgut α-amylase from larvae of A. diaperinus were characterized, and its in vitro activity to proteinaceous preparations from different cultivars of common bean (Phaseolus vulgaris) was determined, as well as the amylolitic activity of insects reared on different types of poultry diet. In order to establish some assay conditions, time course and enzyme concentration upon the reaction rate were determined. Product proceeded linearly with time, and the activity was directly proportional to the enzyme concentration. Banding patterns in mildly denaturing electrophoresis showed a single band with apparent molecular weight of 42 kDa. α-Amylase reached optimal temperature at 45°C and pH 5.0 as the optimal one. It maintained 34.6% of the activity after being kept at 60°C for 5 min, and 23%, after 60 min. However, at 80°C, only 14 and 6% remained after 5 and 60 min, respectively. The presence of Ca and Na ions decreased the enzyme activity at concentrations higher than 2 and 100 mM, respectively. The activity was significantly inhibited by some proteinaceous extracts from common bean cultivars, and it declined with increasing proteinaceous concentration. No significant difference was observed when the amylolytic activity was determined in A. diaperinus reared on different poultry diets, offered to broilers in the starter, grower, finisher, and layer phases.

摘要

小黄粉虫(Alphitobius diaperinus (Panzer))是家禽业的主要害虫,对生产造成严重损害。在本研究中,对小黄粉虫幼虫中肠α-淀粉酶的特性进行了表征,测定了其对不同菜豆(Phaseolus vulgaris)品种蛋白质制剂的体外活性,以及在不同类型家禽日粮饲养下昆虫的淀粉分解活性。为了确定一些测定条件,测定了反应速率随时间和酶浓度的变化。产物随时间呈线性变化,活性与酶浓度成正比。轻度变性电泳中的条带模式显示出一条表观分子量为42 kDa的单带。α-淀粉酶在45°C和pH 5.0时达到最佳温度。在60°C下保持5分钟后,它保持了34.6%的活性,60分钟后保持了23%的活性。然而,在80°C下,5分钟和60分钟后分别仅剩下14%和6%。Ca和Na离子的存在分别在高于2 mM和100 mM的浓度下降低了酶活性。一些菜豆品种的蛋白质提取物显著抑制了该活性,并且活性随蛋白质浓度的增加而下降。在雏鸡、生长鸡、育肥鸡和产蛋鸡阶段给肉鸡饲喂不同家禽日粮饲养的小黄粉虫,测定其淀粉分解活性时未观察到显著差异。

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