Sequential extraction and quantitative recovery of gliadins, glutenins, and other proteins from small samples of wheat flour
Sequential extraction and quantitative recovery of gliadins, glutenins, and other proteins from small samples of wheat flour
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DOI：
10.1021/jf048697l
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发表时间：
2005-03-09
影响因子：
6.1
通讯作者：
Vensel, WH
中科院分区：
文献类型：
--
作者：
DuPont, FM;Chan, R;Vensel, WH
评估了顺序提取和破碎小麦面粉蛋白质的方法，以可靠地定量单个面粉样品中的醇溶蛋白、谷蛋白和白蛋白/球蛋白。通过RP-HPLC结合SIDS-PAGE分析所得蛋白级分的组成。通过质谱或N-末端测序鉴定未知蛋白质。从同一面粉样品中分离和回收离散的白蛋白/球蛋白、麦醇溶蛋白和麦谷蛋白级分的最佳方法是用0.3 M NaI的7.5% 1-丙醇溶液提取，然后用2%SIDS、25 mM DTT的25 mM TRIS溶液（pH 8.0）提取，并用乙酸铵/甲醇沉淀溶解的蛋白质，然后用丙酮沉淀。Butte 86品种的平均面粉成分为10%白蛋白/球蛋白，40%醇溶蛋白和48%谷蛋白。该方法可用于确定不同样品中的面粉成分，并评估蛋白质与最终用途功能之间的关系。
Methods to sequentially extract and fractionate wheat flour proteins were evaluated to reliably quantify gliadins, glutenins, and albumins/globulins in single flour samples. Compositions of the resulting protein fractions were analyzed by RP-HPLC combined with SIDS-PAGE. Unknown proteins were identified by mass spectrometry or N-terminal sequencing. The best separation and recovery of discrete albumin/globulin, gliadin, and glutenin fractions from the same flour sample was achieved by extraction with 0.3 M Nal in 7.5% 1-propanol followed by 2% SIDS, 25 mM DTT in 25 mM TRIS, pH 8.0, and precipitation of the solubilized proteins with ammonium acetate/methanol followed by acetone. Average flour composition for the variety Butte86 was 10% album in/globulin, 40% gliadin, and 48% glutenin. This method should be useful for determining flour composition in diverse samples and evaluating relationships between proteins and end-use functionality.