Purification of wheat flour high-molecular-weight glutenin subunits by fast protein liquid chromatography |
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| Institution: | 1. College of Life Sciences, Northwest A&F University, Yangling Shaanxi 712100, China;2. Department of Biology, Saint Mary''s University, Halifax, Nova Scotia, Canada;3. Key Laboratory of Agricultural Molecular Biology of Shaanxi Province, Yangling Shaanxi 712100, China;1. College of Mechanical and Electronic Engineering, Northwest A&F University, Yangling, Shaanxi 712100, China;2. Department of Biological Systems Engineering, Washington State University, Pullman, WA 99164-6120, USA;1. Institut National de la Santé et de la Recherche Médicale (INSERM) Unit 1033, University of Lyon, Lyon, France;2. Department of Biostatistics, University of Lyon, UMR CNRS 5558, Lyon, France;3. Société de Secours Minière de Bourgogne, Montceau les Mines, France;1. Department of Hematology, Affiliated Hospital of Nantong University, Nantong University, Nantong 226001, Jiangsu Province, People?s Republic of China;2. Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, Nantong 226001, Jiangsu Province, People?s Republic of China;3. Department of Oncology, Affiliated Cancer Hospital of Nantong University, Nantong 226361, Jiangsu Province, People?s Republic of China;1. College of Mechanical and Electronic Engineering, Northwest A&F University, Yangling, Shaanxi 712100, China;2. Department of Food Science and Agricultural Chemistry, McGill University, Montreal H9X 3V9, Canada;3. Department of Biological Systems Engineering, Washington State University, 213 L.J. Smith Hall, Pullman, WA 99164-6120, USA |
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| Abstract: | Fast protein liquid chromatography has been developed for purification of high-molecular-weight glutenin subunits HMW-GSs from wheat flour. Flour samples from four wheat cultivars with different HMW-GS alleles at Glu-A1, Glu-B1 and Glu-D1 loci were used to establish the method. The column material used was Resource? Phe, and the optimal elution was with a gradient formed with buffer A 0.05 M Tris–HCl containing 4 M urea and 0.25 M (NH4)2SO4, pH 8.0] and buffer B 0.05 M Tris–HCl containing 4 M urea (pH 8.0)] at a flow rate of 0.5 ml/min. A pure single 1Dx-, 1Bx- HMW-GS, and all the y-type HMW-GSs present in one genotype can be reliably separated in a single step. |
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