Kinetics, aggregation behavior and optimization of the fractionation of whey protein isolate with hydrochloric acid☆

Abstract Concentrated WPI solutions (10%, w/w) containing approximately 50% beta-lactoglobulin (β-LG) and 26% alpha-lactalbumin (α-LA) were fractionated with HCl at acidic pH and moderate temperature to obtain enriched α-LA and β-LG fractions. Aggregation behavior and kinetics of protein precipitation and aggregate formation were analyzed as a function of four process parameters: pH (3.0–5.5), temperature (50–70 °C), reaction time (0–180 min) and protein concentration (10–29%). The precipitation and aggregation of α-LA appeared rate-limited, with a logarithmic dependence of time and possible bimodal nucleation rate, and varied considerably with pH and temperature. Aggregates as large as ∼300 μm were noted after 120 min at pH 4, 60 °C. Processing parameters were optimized to obtain both a high aggregate yield and optimal composition of the aggregate fraction. The optimally enriched solid and liquid fractions contained 58% α-LA and 76% β-LG, respectively, with 99% and 74% recovery ratios. Over the pH range studied, β-LG aggregation was found negligible at 60 °C and β-LG recovery in the aggregates attributed to liquid holding. Increasing WPI concentration accelerated α-LA aggregation, demonstrating a concentration-dependent aggregation mechanism, and reduced aggregate purity. Enriched whey protein fractions are valuable health-enhancing food ingredients.