Stability of wheat proteins in solution

Gluten from defatted wheat flour was used to study the stability of protein extracts during various steps that are routinely made in protein analyses. These included aqueous extraction and short-term storage, handling of chromatographic fractions, freeze-drying and reconstitution. Protein composition was monitored by size-exclusion high performance liquid chromatography (SE-HPLC) and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Distilled, deionized and sterilized water were used to prepare the gluten solutions (TFA or HCl solutions at pH 3.0) and to prepare elution solvents. In distilled or deionized water there was a decrease of polymeric protein and a corresponding increase in the monomeric components with time unless initial heating was applied. Protein extracts obtained from gluten and solvents prepared with freshly sterilized water showed no change during storage for 4 days at a temperature of 25 °C. Microbiological assays of the water samples supported the view that microbial contaminants were the source of proteases. After elution from SE-HPLC column with acetonitrile/water followed by freeze-drying and re-dissolution, the SE-HPLC profile of the monomeric proteins was unchanged. However, polymeric proteins aggregated, lost solubility and showed an altered SE-HPLC profile. The effect appeared to be due to the presence of acetonitrile and could be avoided by membrane concentration instead of freeze-drying.