Meireles Elaine A, Carneiro Cíntia N B, DaMatta Renato A, Samuels Richard I, Silva Carlos P
Laboratório de Químnica e Função de Proteínas e Peptídeos, Universidade Estadual do Norte Fluminense Darcy Ribeiro, CEP 28013-620, Campos dos Goytacazes, RJ, Brasil.
J Insect Sci. 2009;9:43. doi: 10.1673/031.009.4301.
Scanning electron microscopy images were taken of starch granules from different sources following exposure in vivo and in vitro to gut alpha-amylases isolated from Tenebrio molitor L. (Coleoptera: Tenebrionidae) and Zabrotes subfasciatus Boheman (Coleoptera: Bruchidae). One alpha-amylase was isolated from whole larval midguts of T. molitor using non-denaturing SDS-PAGE, while two other alpha-amylase fractions were isolated from whole larval midguts of Z. subfasciatus using hydrophobic interaction chromatography., Digested starch granules from larvae fed on maize, potato or wheat were isolated from midgut contents. Combinations of starch granules with isolated alpha-amylases from both species showed similar patterns of granule degradation. In vitro enzymatic degradation of maize starch granules by the three different alpha-amylase fractions began by creating small holes and crater-like areas on the surface of the granules. Over time, these holes increased in number and area resulting in extensive degradation of the granule structure. Granules from potato did not show formation of pits and craters on their surface, but presented extensive erosion in their interior. For all types of starch, as soon as the interior of the starch granule was reached, the inner layers of amylose and amylopectin were differentially hydrolyzed, resulting in a striated pattern. These data support the hypothesis that the pattern of starch degradation depends more on the granule type than on the alpha-amylase involved.
对来自不同来源的淀粉颗粒进行扫描电子显微镜成像,这些淀粉颗粒在体内和体外暴露于从黄粉虫(鞘翅目:拟步甲科)和赤拟谷盗(鞘翅目:豆象科)分离的肠道α-淀粉酶。使用非变性SDS-PAGE从黄粉虫的整个幼虫中肠分离出一种α-淀粉酶,而使用疏水相互作用色谱法从赤拟谷盗的整个幼虫中肠分离出另外两种α-淀粉酶组分。从取食玉米、马铃薯或小麦的幼虫中肠内容物中分离出消化后的淀粉颗粒。淀粉颗粒与来自这两个物种的分离α-淀粉酶的组合显示出相似的颗粒降解模式。三种不同的α-淀粉酶组分对玉米淀粉颗粒的体外酶促降解开始于在颗粒表面形成小孔和火山口状区域。随着时间的推移,这些孔的数量和面积增加,导致颗粒结构的广泛降解。马铃薯颗粒表面未显示出凹坑和火山口的形成,但其内部出现广泛侵蚀。对于所有类型的淀粉,一旦到达淀粉颗粒内部,直链淀粉和支链淀粉的内层就会被不同程度地水解,形成条纹状图案。这些数据支持了淀粉降解模式更多地取决于颗粒类型而非所涉及的α-淀粉酶这一假设。