A comparative study of physicochemical and conformational properties in three vicilins from Phaseolus legumes: Implications for the structure–function relationship

The polypeptide constituents, physicochemical and conformational properties of three vicilins from kidney, red and mung beans, fractionated using combined acid extraction and ion-exchange chromatographic techniques, were characterized and compared. The tested physicochemical and conformational properties included amino acid composition, surface charge and hydrophobicity, protein solubility (PS; as a function of pH), emulsifying activity index (EAI) and emulsion stability index (ESI), as well as secondary, tertiary and/or quaternary conformations. The amino acid composition (especially relative acidic/basic charged amino acid ratio, and the percentage of polar uncharged amino acids) was an intrinsic physicochemical parameter that largely determined other physicochemical and conformational properties. The PS and EAI (and ESI) of these vicilins at various pH values were associated with each other, and both were closely related to their surface charge and/or hydrophobicity. Far-UV CD spectral analyses indicated that these vicilins exhibited similar secondary structure compositions at pH deviating from the isoelectric point (about pH 4.5). However, their tertiary and/or quaternary conformations, as determined by differential scanning calorimetry, intrinsic fluorescence and near-UV CD spectroscopy, remarkably varied. At pH 7.0 or 9.0, there were close relationships between the EAI and the flexibility in quaternary conformation, and between the ESI and the flexibility in tertiary conformation. This is the first observation to elucidate the importance of tertiary and/or quaternary conformations for the emulsifying properties of legume vicilins. Additionally, an underlying mechanism of the emulsification process for the vicilins (trimeric form) with excellent emulsifying properties was proposed.