Characterization of HMW Glutenin Subunits in Bread and Tetraploid Wheats by Reversed-Phase High-Performance Liquid Chromatography
首发时间:2009-01-15
Abstract:The wheat storage proteins, especially the high molecular weight glutenin subunits (HMW-GS), play important roles in the determination of flour processing and bread-making quality. Compared with the traditional SDS-PAGE method, reversed-phase high-performance liquid chromatography (RP-HPLC) was shown to have many advantages for the separation and characterization of HMW-GS because of its high resolving power, repeatability and automation. In this work, HMW-GS from bread and tetraploid wheats were separated and characterized by RP-HPLC. The elution time ranking of different HMW-GS was: 1Ax>1Bx>1Dx>1By>1Dy. Several subunit pairs associated with good quality properties and those with similar mobilities on SDS-PAGE, such as 1Bx7 and 1Bx7*, 1By8 and 1By8*, 1Dx2 and 1Ax2*, 1Bx6 and 1Bx6.1, were well separated and readily identified through RP-HPLC. However, other subunit pairs, such as 1Dy10-1Dy12, 1Dx5-1By18 and 1Dx2-1By16, could not be adequately separated and identified by RP-HPLC, whereas they displayed different mobilities on SDS-PAGE gels. Because 1Dx5 and 1Dx2 showed different hydrophobicities, RP-HPLC could distinguish 1Dx5+1Dy10 and 1Dx2+1Dy12. A comparative analysis between RP-HPLC and SDS-PAGE showed that a combination of both methods provided more effective identification of HMW-GS in wheat quality improvement and germplasm screening.
keywords: Wheat HMW-GS RP-HPLC
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应用反相高效液相色谱技术分离小麦高分子量谷蛋白亚基
摘要:小麦高分子量谷蛋白亚基(HMW-GS)是面包小麦重要的贮藏蛋白,决定了面团的粘弹性,与面包的烘烤品质密切相关。与传统的SDS-PAGE方法相比,反相高效液相色谱(RP-HPLC)在分析分离贮藏蛋白的过程中,具有高溶解性、可重复性、自动控制等优点。本文应用反相高效液相色谱技术分离分析普通面包小麦和四倍体小麦后发现,根据洗脱时间的不同,高分子量谷蛋白亚基的出峰顺序依次为:1Ax>1Bx>1Dx>1By>1Dy。在SDS-PAGE中,一些与优质品质相关的亚基,如1Bx17和1By18,1Bx20x和1By20y, 1Bx7和1Bx7*, 1By8和1By8*, 1Dx2和1Ax2*,具有相同或相似的迁移率,彼此较难区分,但是通过RP-HPLC技术可以获得很好的分离效果。同时,一些亚基对,如1Dy10-1Dy12, 1Dx5-1By18和1Dx2-1By16,可以通过SDS-PAGE获得很好的分离,而通过RP-HPLC的分离效果却不甚理想。但是由于1Dx5和1Dx2具有不同的疏水性,通过RP-HPLC 技术可以很好的区分1Dx5+1Dy10和1Dx2+1Dy12两组亚基对。本文通过比较传统的SDS-PAGE方法和RP-HPLC技术认为,将两种方法有效地结合起来分离分析小麦高分子量谷蛋白亚基,对进一步揭示小麦高分子量谷蛋白亚基与面包小麦烘烤品质之间的关系,提供了有力的技术支持。
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No.2786739049012320****
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