Simultaneous Isolation of Wheat High Molecular Weight and Low Molecular Weight Glutenin Subunits |
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| Affiliation: | 1. Institute of Occupational Medicine and Maritime Medicine (ZfAM), University Medical Center Hamburg-Eppendorf, Hamburg, Germany;2. Practice Müsken, Bad Lippspringe, Germany;3. Biocenter Klein Flottbek and Botanical Garden, University of Hamburg, Hamburg, Germany;3. From the Allergy Division, Department of Medicine, University of Virginia, Charlottesville, Virginia 22903,;4. INDOOR Biotechnologies, Inc., Charlottesville, Virginia 22908,;5. the Joint School of Nanoscience and Nanoengineering, University of North Carolina, Greensboro, North Carolina 27401,;6. the Macromolecular Crystallography Laboratory, National Cancer Institute, National Institutes of Health, Frederick, Maryland 21702,;12. the Department of Molecular Biology, University of Salzburg, 5020 Salzburg, Austria;1. Department of Chemistry, University of Missouri-Columbia, Columbia, MO 65211, USA;2. Veterinary Medical Diagnostic Laboratory, University of Missouri-Columbia, Columbia, MO 65211, USA;3. Department of Veterinary Pathobiology, University of Missouri-Columbia, Columbia, MO 65211, USA;4. Department of Biochemistry, University of Missouri-Columbia, Columbia, MO 65211, USA;1. Institute of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria;2. Molecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook University, Townsville, QLD, Australia;3. Australian Institute of Tropical Health and Medicine, James Cook University, Townsville, QLD, Australia;4. Center for Molecular Therapeutics, James Cook University, Townsville, QLD, Australia;5. School of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, VIC, Australia;6. Department of Paediatrics, Murdoch Children''s Research Institute, The University of Melbourne, Flemington Road, Parkville, VIC, Australia;7. VetCore Facility for Research, University of Veterinary Medicine, Vienna, Austria;1. Centro de Investigação em Química da Universidade do Porto (CIQUP), Faculdade de Ciências da Universidade do Porto, Departamento de Química e Bioquímica, Rua do Campo Alegre 687, 4169-007, Porto, Portugal;2. REQUIMTE-LAQV/Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, Porto, Portugal |
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| Abstract: | A procedure was established for isolating simultaneously high molecular weight (HMW-GS) and low molecular weight glutenin subunits (LMW-GS) from chloroform-defatted wheat flour. Temperature conditions were optimised to obtain maximum yield and purity. A pre-extraction of gliadin was performed with 50% (v/v) propan-1-ol at room temperature. Glutenin subunits were solubilised from the residue at 60 °C with 50% (v/v) propan-1-ol containing 1% (w/v) dithiothreitol. The HMW-GS in the glutenin mixture were then precipitated selectively by increasing the propan-1-ol concentration to 60%. Upon addition of propan-1-ol to a final concentration of 85%, LMW-GS were precipitated. It was shown by densitometric scanning of SDS–PAGE gels that the isolated glutenin subunits were very pure. Furthermore, a partial separation of LMW-GS into B and C type subunits was observed when the propan-1-ol concentration was increased gradually. |
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