Molecular changes of phenolic-protein interactions in isolated proteins from flaxseed and soybean using Native-PAGE, SDS-PAGE, RP-HPLC, and ESI-MS analysis.

Abstract

The effects of protein-phenolic interactions on the molecular characteristics of soybean and flaxseed proteins were investigated. Proteins were isolated from soybean and flaxseed using isoelectric precipitation, followed by extraction of free and bound phenolics. The effects of elimination of the phenolic compounds on molecular characteristics of the protein isolates were studied using reversed phase-high performance liquid chromatography (RP-HPLC), Native and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and electron spray ionization-mass spectrometry (ESI-MS). The Native-PAGE fractions from isolated proteins from full-fat flaxseed and soybean revealed that protein migration was affected by removal of bound phenolics. SDS-PAGE from full-fat and defatted protein isolates of flaxseed and soybean revealed that the removal of bound phenolics affected the molecular characteristics of protein subunits. Soybean protein isolates had protein-phenolic interactions through acidic and basic subunits. RP-HPLC and ESI-MS showed that the removal of bound and free phenolics had only minor effects on the molecular characteristics of isolated proteins from defatted and full-fat soybean. With respect to isolated proteins from flaxseed, the removal of bound phenolics showed little effect on the electrophoretic behavior of the proteins or the protein subunits. PRACTICAL APPLICATIONS: Phenolic-protein and phenolic-lipid-protein interactions may affect the nutritional, physicochemical, and functional properties of isolated proteins from food in flaxseed and soybean.