Xu H, Wang R J, Shen X, Zhao Y L, Sun G L, Zhao H X, Guo A G
College of Life Sciences, Northwest A & F University, Yangling, Shaanxi, 712100, People's Republic of China.
Theor Appl Genet. 2006 Nov;113(7):1295-303. doi: 10.1007/s00122-006-0383-2. Epub 2006 Aug 24.
Some allelic forms of low-molecular-weight glutenin subunit (LMW-GS) can greatly influence the end-use of wheat flours, understanding the function of each allele of LMW-GS is important to wheat quality breeding. A LMW-GS gene XYGluD3-LMWGS 1(AY263369) has been cloned from bread wheat cultivar Xiaoyan 6. The deduced protein contained nine cystine residues, one more than that in all other LMW-GSs reported previously, indicating that it is either a new gene or a new allele of a known LMW-GS gene. In this study, the gene was expressed in E. coil in large scale for the testing of its functional property. Reactive Red 120-Agarose resin was used efficiently to purify the expressed LMW-GS proteins from bacteria, with the lactic acid-sodium lactate buffer (pH 4.5) which contained low concentration SDS as elution solution. The purified protein (belonging to the LMW-m family, MW about 35 KDa) was supplemented into a base flour, the results of 10 g dough mixing test indicated that incorporation of the LMW-GS increased the strength of the dough, with significant increases in mixing time (MT) and peak width (PW), and decrease in breakdown in resistance (RBD) compared with the control. In addition, the dough with incorporation of the LMW-GS had more glutenin macropolyeric protein than the control, suggesting that the LMW-GS participated in forming larger glutenin polymers, and greatly contributed to dough strength. The changes in mixing parameters and the amount of glutenin macropolyeric protein were related to the quantity of incorporating subunits.
一些低分子量麦谷蛋白亚基(LMW-GS)的等位基因形式会极大地影响小麦粉的最终用途,了解LMW-GS每个等位基因的功能对于小麦品质育种很重要。已从小麦品种小偃6中克隆出一个LMW-GS基因XYGluD3-LMWGS 1(AY263369)。推导的蛋白质含有9个半胱氨酸残基,比先前报道的所有其他LMW-GS多一个,表明它要么是一个新基因,要么是已知LMW-GS基因的一个新等位基因。在本研究中,该基因在大肠杆菌中大规模表达以测试其功能特性。使用活性红120-琼脂糖树脂有效地从细菌中纯化表达的LMW-GS蛋白,以含有低浓度SDS的乳酸-乳酸钠缓冲液(pH 4.5)作为洗脱液。将纯化的蛋白(属于LMW-m家族,分子量约35 kDa)添加到基础面粉中,10 g面团混合试验结果表明,与对照相比,添加LMW-GS提高了面团的强度,混合时间(MT)和峰值宽度(PW)显著增加,抗阻力衰减(RBD)降低。此外,添加LMW-GS的面团比对照含有更多的麦谷蛋白大聚合物蛋白,表明LMW-GS参与形成更大的麦谷蛋白聚合物,并对面团强度有很大贡献。混合参数和麦谷蛋白大聚合物蛋白量的变化与添加亚基的量有关。