Extraction of up to 95% of wheat (Triticum aestivum) flour protein using warm sodium dodecyl sulfate (SDS) without reduction or sonication.
Extraction of glutenin polymers without sonication is an essential prerequisite for accurate determination of their composition and molecular size distribution. Sequential fractionation of wheat flour with 0.1 M KCl and 0.25% sodium dodecyl sulfate (SDS) at 21 degrees C and 2% SDS at 60 degrees C extracted up to 95% of total protein. We propose that 2% SDS at 60 degrees C disrupts hydrogen bonds in glutenin and gliadin aggregates, reduces hydrophobic interactions, and facilitates solubilization. Analysis by size-exclusion high-performance liquid chromatography (SE-HPLC), reverse-phase (RP)-HPLC, and SDS-polyacrylamide gel electrophoresis (PAGE) revealed that partitioning of gliadins and glutenins among the extracts differed for two flours with good baking quality (Butte 86 and Jagger) and one with poor baking quality (Chinese Spring). More gliadin was associated with the 0.25% SDS extract for Chinese Spring, whereas more gliadin was associated with the 2% SDS extract for Butte 86 and Jagger. Unextractable glutenin polymer was only 4-5% of total protein for Butte 86 and Chinese Spring and 14% for Jagger.
DuPont
Frances M
FM
United States Department of Agriculture, Agricultural Research Service, Western Regional Research Center, 800 Buchanan Street, Albany, California 94710, USA. frances.dupont@ars.usda.gov
Samoil
Vitalie
V
Chan
Ronald
R
D005591:Chemical Fractionation; D005433:Flour / Q000032:analysis*; D005903:Gliadin / Q000302:isolation & purification; D005983:Glutens / Q000302:isolation & purification; D006358:Hot Temperature; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification*; D011189:Potassium Chloride; D012967:Sodium Dodecyl Sulfate; D013010:Sonication; D014908:Triticum / Q000737:chemistry*
D010940:Plant Proteins; D012967:Sodium Dodecyl Sulfate; D011189:Potassium Chloride; D005983:Glutens; D005903:Gliadin; C037302:glutenin