Silva C P, Terra W R, Xavier-Filho J, Grossi de Sá M F, Isejima E M, DaMatta R A, Miguens F C, Bifano T D
Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, CEP 28015-620, Campos dos Goytacazes, Brazil.
Insect Biochem Mol Biol. 2001 Jan;31(1):41-50. doi: 10.1016/s0965-1748(00)00103-x.
Zabrotes subfasciatus larvae possess three alpha-amylase isoforms as determined by in gel assays following SDS-PAGE. The two minor isoforms present lower electrophoretic mobility than the major form, and seem to occur as a heterodimer. When developed inside Vigna unguiculata (cowpea) seeds, fourth instar larvae have minor quantities of the slow-migrating forms, but when reared on seeds of Phaseolus vulgaris (common bean) or Phaseolus lunatus, the two slow-migrating forms are expressed in higher amounts, while activity of the major form was independent of the host seed. Larvae developing inside cowpea seeds at the beginning of the fourth instar were fed on flour from cotyledons of cowpea or common bean. Larvae fed on the common bean flour started to express the dimer in higher amounts when compared with the control larvae fed on cowpea flour. In an attempt to correlate differences between starch granules and the induction of alpha-amylases, a detailed study on the digestive process of the granules was conducted. Incorporation of purified starch granules into artificial diets did not induce the two minor alpha-amylases. The in vitro hydrolysis rates of purified granules and the pattern of dextrins liberated by the different alpha-amylases were similar for the two legume species. The starch granules enter the midgut extensively damaged, which may facilitate the access to the more susceptible parts of the granules to enzymatic attack.
通过SDS-PAGE后的凝胶分析确定,豆象幼虫拥有三种α-淀粉酶同工型。两种次要同工型的电泳迁移率低于主要同工型,且似乎以异二聚体形式存在。在豇豆种子内发育时,四龄幼虫体内的慢速迁移形式含量较少,但在菜豆或利马豆种子上饲养时,两种慢速迁移形式的表达量更高,而主要同工型的活性与宿主种子无关。在四龄初期于豇豆种子内发育的幼虫,以豇豆或菜豆子叶的面粉为食。与以豇豆面粉为食的对照幼虫相比,以菜豆面粉为食的幼虫开始大量表达二聚体。为了关联淀粉颗粒与α-淀粉酶诱导之间的差异,对颗粒的消化过程进行了详细研究。将纯化的淀粉颗粒掺入人工饲料中不会诱导两种次要的α-淀粉酶。两种豆科植物的纯化颗粒的体外水解速率以及不同α-淀粉酶释放的糊精模式相似。淀粉颗粒进入中肠时受到广泛破坏,这可能有助于酶更容易地攻击颗粒中更易受影响的部分。