College of Life Science, Capital Normal University , 100048 Beijing, China.
J Agric Food Chem. 2013 May 1;61(17):4026-34. doi: 10.1021/jf400472s. Epub 2013 Apr 17.
Low molecular weight glutenin subunits (LMW-GS), as important seed storage proteins, together with HMW-GS significantly define unique dough viscoelastic properties. In this study, a rapid ultraperformance liquid chromatography (UPLC) method for the separation and characterization of LMW-GS in wheat was optimized and established. The fast, reproducible, and high-resolution UPLC separation of LMW-GS could be obtained by gradually increasing eluting gradient from 21 to 47% in 30 min at flow rate of 0.55 mL/min and 60 °C for separation temperature. By this method, analysis of one sample could be completed in <20 min, significantly less time than the traditional reversed-phase high-performance liquid chromatography (RP-HPLC) method. Under the optimized conditions, the genetic features of LMW-GS and genotype × environmental interaction were successfully analyzed, leading to a fast identification of 17 main LMW-GS alleles that were related to different quality properties in wheat. The results demonstrated that UPLC could be a powerful and alternative tool for genetic and proteomic studies of wheat grain proteins and fast identification or screening of desirable LMW-GS alleles in wheat quality improvement.
低分子量谷蛋白亚基(LMW-GS)作为重要的种子贮藏蛋白,与高分子量谷蛋白亚基(HMW-GS)一起显著定义了独特的面团黏弹性。在本研究中,优化并建立了一种快速超高效液相色谱(UPLC)法用于小麦 LMW-GS 的分离和鉴定。通过在 30 分钟内逐渐将洗脱梯度从 21%增加到 47%,在流速为 0.55 mL/min 和 60°C 的分离温度下,可实现 LMW-GS 的快速、可重现和高分辨率 UPLC 分离。通过该方法,<20 分钟即可完成一个样品的分析,明显快于传统反相高效液相色谱(RP-HPLC)法。在优化条件下,成功分析了 LMW-GS 的遗传特征和基因型×环境互作,快速鉴定了与小麦不同品质特性相关的 17 个主要 LMW-GS 等位基因。结果表明,UPLC 可以成为小麦籽粒蛋白质遗传和蛋白质组学研究的有力工具,也可以快速鉴定或筛选小麦品质改良中所需的 LMW-GS 等位基因。