Expression, Purification, and Functional Analysis of Three Low‐Molecular‐Weight Glutenin Subunits from Wheat Cultivar Cheyenne

Abstract

Cereal Chem. 91(4):378-382Glutenins, which form the network of gluten protein, are of great importance for the quality of flour products. Glutenins can be divided into HMW and LMW subunits according to molecular weight. Three genes for LMW glutenin subunits (LMW-GS), named lmw-cnd1, lmw-cnd2, and lmw-cnd3 with open reading frames of 1,053, 903, and 969 bp, respectively, were cloned from wheat cultivar Cheyenne. Heterologous expression vectors of the three LMW-GS were constructed, and the recombinant proteins LMW-CND1, LMW-CND2, and LMW-CND3 were overexpressed in Escherichia coli. After cell disruption with ultrasound, target proteins of high purity were obtained by using Ni 2+ affin-ity chromatography. Farinograph and TAPlus measurements were used to investigate the effects of the three LMW-GS on the characteristics of flour and dough. The results showed that the addition of each LMW-GS can lead to an increase in the elasticity of the dough. Moreover, LMW-CND2 and LMW-CND3 promoted the strength of the dough. All three LMW-GS caused a decrease of hardness and increase of springiness and cohesiveness of dough according to texture profiling results. Consequently, all three LMW-GS have positive effects on the processing characteristics of dough and can improve bread quality to different extents.* The e-Xtra logo stands for "electronic extra" and indicates that Figures 1 and 2 appear in color online.