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王珏, 罗立廷, 陈明洁, 李洋, 张金锐, 何光源. 小麦面粉Puroindoline蛋白的提取与纯化[J]. 植物科学学报, 2005, 23(6): 568-571.
引用本文: 王珏, 罗立廷, 陈明洁, 李洋, 张金锐, 何光源. 小麦面粉Puroindoline蛋白的提取与纯化[J]. 植物科学学报, 2005, 23(6): 568-571.
WANG Jue, LUO Li-Ting, CHEN Ming-Jie, LI Yang, ZHANG Jin-Rui, HE Guang-Yuan. Extraction and Purification of Puroindoline from Wheat Flour[J]. Plant Science Journal, 2005, 23(6): 568-571.
Citation: WANG Jue, LUO Li-Ting, CHEN Ming-Jie, LI Yang, ZHANG Jin-Rui, HE Guang-Yuan. Extraction and Purification of Puroindoline from Wheat Flour[J]. Plant Science Journal, 2005, 23(6): 568-571.

小麦面粉Puroindoline蛋白的提取与纯化

Extraction and Purification of Puroindoline from Wheat Flour

  • 摘要: Puroindoline蛋白是小麦面粉中一种非常重要的蛋白质,不仅影响和决定了籽粒的硬度,而且有抗G+、G-菌以及抗真菌的作用。用含4% Triton X-114、100mmol/L pH7.8Tris-HCl缓冲液处理小麦面粉来分离Puroindo-line蛋白,经处理后得到的蛋白质混合溶液首先用分子筛葡聚糖G-75纯化,每个收集管内的组分经SDS-PAGE分析,分子量小于31kD的蛋白质组分被回收和集中,回收的蛋白质组分经PEG20000浓缩后,再用离子交换柱羧甲基纤维素(CM-23)进行纯化。其洗脱液分别是双蒸水和NaCl,梯度为0.05~0.7mol/L、8mmol/L pH5.5的MES缓冲液,回收只含15kD的蛋白质的组分,接着用PEG20000浓缩,最后冷冻干燥得到Puroindoline蛋白。

     

    Abstract: Puroindoline,influenced and decided the wheat hardness and exhibiting activity against gram-positive bacteria,gram-negative bacteria and fungi,is a very important protein in wheat flour.In this studies,the puroindoline was extracted from wheat flour in the 100 mmol/L Tris-HCl pH7.8 buffer,containing 4% Triton X-114.Then the protein mixture including puroindoline was successively purified by Sephadex G-75,and each fraction was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Fractions containing polypeptides with apparent molecular weights of less than 31 kD were pooled and loading on Carboxymethyl-cellulose(CM-23) after concentrated by PEG20000,and each fraction was analyzed as preceding.In those ways,they were eluted by distilled water and applying a gradient from 0.05 mol/L to(0.7 mol/L) NaCl in 8 mmol/L MES pH5.5 buffer respectively.The fractions containing polypeptides apparent molecular weight 15 kD were pooled and concentrated by PEG20000,at last the puroindoline was obtained by freeze-dried.

     

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