Kinetics of Formation and Functional Properties of Conjugates Prepared by Dry-State Incubation of β-Lactoglobulin/Acacia Gum Electrostatic Complexes

Abstract

The formation of conjugates between beta-lactoglobulin and acacia gum based on electrostatic complexes formed at pH 4.2 was investigated upon dry-state incubation for up to 14 days at 60 degrees C and 79% relative humidity (RH). By means of SEC-HPLC and RP-HPLC, it was shown that the beta-lactoglobulin incubated alone was able to form polymers with molecular masses higher than 200 kDa until 50% of the initial monomeric protein disappeared after 14 days. In the presence of acacia gum at initial protein to polysaccharide weight mixing ratios of 2:1 and 1:2, only 35% of the initial beta-lactoglobulin monomers disappeared after 14 days. Using RP-HPLC, an apparent reaction order of 2 was found for the disappearance of monomeric beta-lactoglobulin both in the presence or absence of acacia gum. However, the reaction rate was faster in the absence of acacia gum. SDS-PAGE electrophoresis with silver staining confirmed the formation of beta-lactoglobulin/acacia gum conjugates. The solubility curves of the incubated beta-lactoglobulin showed a minimum around pH 4-5. By contrast, the minimum of solubility of the beta-lactoglobulin/acacia gum incubated mixtures shifted to lower pH values compared to initial mixtures. The conjugates exhibited higher foam capacity than the incubated protein as well as lower equilibrium air/water surface tension. Conjugation at ratio 1:2 led to increased interfacial viscosity (300 mN s m(-1) at 0.01 Hz) compared to beta-lactoglobulin alone (100 mN s m(-1) at 0.01 Hz), but similar interfacial elasticity (30-40 mN m(-1)). The foam capacity of the conjugates was significantly higher than that of the incubated beta-lactoglobulin as well as foam expansion and drainage time, especially at pH 5.3, i.e., higher than the pH of formation of the conjugates.