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Extraction and purification of low molecular weight glutenin subunits using size exclusion chromatography

  • Priya Dangi
  • B. S. KhatkarEmail author
Original Article
  • 56 Downloads

Abstract

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 (Mw 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 (Mw < 35 kDa) were eluted in peak III. SEC proved to be a valuable tool in purifying LMW-GS in a functionally active state.

Keywords

Size-exclusion chromatography LMW-GS SDS-PAGE 

Notes

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Copyright information

© Association of Food Scientists & Technologists (India) 2019

Authors and Affiliations

  1. 1.Department of Food TechnologyGuru Jambheshwar University of Science and TechnologyHisarIndia

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