Departamento de Biologia Vegetal, Instituto de Biologia, Universidade Estadual de Campinas, CP 6109, 13083-970, Campinas, SP, Brazil.
J Agric Food Chem. 2010 Mar 10;58(5):3050-5. doi: 10.1021/jf9037216.
Legumin-like proteins from seeds of Coffea arabica (CaL-1 and CaL-2) and Coffea racemosa (CrL-1 and CrL-2) were characterized and isolated by gel filtration and reverse-phase chromatography. The insecticidal properties of the purified proteins were tested against Callosobruchus maculatus using artificial diets. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses indicated that CaL-1 is composed of two subunits of 33 and 24 kDa, while CaL-2, CrL-1, and CrL-2 were monomeric with a single band of 14 kDa. The LD(50) values were 0.5% (w/w) for CaL-1 and 0.3% (w/w) for CaL-2, CrL-1, and CrL-2. ED(50) at 0.3% was assessed for all protein concentrations. The legumin-like proteins were not digested by midgut homogenates of C. maculatus until 8 h of incubation. CaL-1 and CaL-2 ( C. arabica ) and CrL-1 and CrL-2 ( C. racemosa ) are chitin-binding proteins, and their insecticidal properties toward C. maculatus larvae might be related to their capacity to bind chitin present in the larval gut and their associated low digestibility.
从阿拉比卡咖啡（CaL-1 和 CaL-2）和罗布斯塔咖啡（CrL-1 和 CrL-2）的种子中分离和鉴定了类豆球蛋白样蛋白，采用凝胶过滤和反相色谱法。利用人工饲料测试了纯化蛋白对绿豆象的杀虫特性。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析表明，CaL-1 由 33 和 24 kDa 的两个亚基组成，而 CaL-2、CrL-1 和 CrL-2 则为单体，只有 14 kDa 的单带。CaL-1 的 LD(50)值为 0.5%（w/w），CaL-2、CrL-1 和 CrL-2 的 LD(50)值为 0.3%（w/w）。在 0.3%的 ED(50)下评估了所有蛋白浓度。类豆球蛋白样蛋白在孵育 8 小时内不会被绿豆象中肠匀浆消化。CaL-1 和 CaL-2（阿拉比卡咖啡）和 CrL-1 和 CrL-2（罗布斯塔咖啡）是几丁质结合蛋白，它们对绿豆象幼虫的杀虫特性可能与其结合幼虫肠道中存在的几丁质的能力及其相关的低消化率有关。
