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Optimization of a protein extraction method from natural rubber sheets made of Hevea brasiliensis latex

机译:HEVEA Brasiliensis乳胶制成的天然橡胶板蛋白质提取方法的优化

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A method to extract proteins from natural rubber (NR) samples has been developed to characterize the protein composition of NR by SDS-PAGE electrophoresis. While the incubation of NR pieces in aqueous buffers did not provide satisfactory results (no protein bands detected by SDS-PAGE), the solubilization of NR pieces in an organic solvent, prior to centrifugation and protein extraction from the obtained pellet, was found efficient to highlight protein bands by SDS-PAGE. Pure cyclohexane was used to solubilize NR pieces and resulted in a low share of nitrogen in the pellet (29%). The addition of 10% (v/v) ethanol to cyclohexane allowed to significantly increase this share to 73% and to detect 13 protein bands by SDS-PAGE. The molecular weights of the four most intense bands suggest the presence in NR of the following proteins: rubber elongation factor, small rubber particle protein, hevamine, and beta-1-3 glucanase.
机译:建立了一种从天然橡胶(NR)样品中提取蛋白质的方法,用SDS-PAGE电泳表征NR的蛋白质组成。虽然NR片段在水性缓冲液中的培养没有提供令人满意的结果(SDS-PAGE未检测到蛋白条带),但在离心和从获得的颗粒中提取蛋白质之前,NR片段在有机溶剂中的增溶被发现可以通过SDS-PAGE有效地突出蛋白条带。纯环己烷用于溶解NR块,导致颗粒中的氮含量较低(29%)。向环己烷中添加10%(v/v)乙醇,可使该比例显著增加至73%,并通过SDS-PAGE检测到13条蛋白质条带。四条最密集条带的分子量表明NR中存在以下蛋白质:橡胶伸长因子、小橡胶颗粒蛋白质、hevamine和β-1-3葡聚糖酶。

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