Dangi Priya, Khatkar B S
Department of Food Technology, Guru Jambheshwar University of Science and Technology, Hisar, Haryana 125001 India.
J Food Sci Technol. 2019 Feb;56(2):951-956. doi: 10.1007/s13197-018-03560-1. Epub 2019 Jan 11.
Crude glutenin of commercial Indian wheat varieties was fractionated into high molecular weight glutenin subunits (HMW-GS) and low molecular weight glutenin subunits (LMW-GS) by employing size-exclusion chromatography (SEC). The SEC profile of glutenins obtained with different buffers were discriminated effectively with respect to the quality of the proteins eluted in each peak. The most efficient separation of LMW-GS was achieved using 3 M urea (pH 5.5) buffer under unreduced conditions. The chromatogram was segregated predominantly into three peaks with varied molecular weights as determined by SDS-PAGE. Peak I corresponded to a mixture of HMW-GS and LMW-GS (M 100-30 kDa). Peak II enclosed LMW-GS specifically with molecular weights in the range of 45-35 kDa. Lastly, a mixture of proteins associated with LMW-GS (M < 35 kDa) were eluted in peak III. SEC proved to be a valuable tool in purifying LMW-GS in a functionally active state.
通过尺寸排阻色谱法（SEC）将印度商业小麦品种的粗谷蛋白分离为高分子量谷蛋白亚基（HMW-GS）和低分子量谷蛋白亚基（LMW-GS）。就每个峰中洗脱的蛋白质质量而言，用不同缓冲液获得的谷蛋白的SEC图谱得到了有效区分。在未还原条件下，使用3M尿素（pH 5.5）缓冲液可实现LMW-GS的最有效分离。通过SDS-PAGE测定，色谱图主要分离为三个分子量不同的峰。峰I对应于HMW-GS和LMW-GS的混合物（M 100-30 kDa）。峰II专门包含分子量在45-35 kDa范围内的LMW-GS。最后，与LMW-GS相关的蛋白质混合物（M < 35 kDa）在峰III中洗脱。SEC被证明是纯化功能活性状态下LMW-GS的有价值工具。
