Amino acid composition and biological value of whey protein hydrolysates fractionated by cascade-selective membrane filtration

This article discusses the application of industrial cascade-selective membrane filtration techniques for the purification of enzymatic hydrolysates of whey proteins and the fractionation of bioactive peptides. The main focus is on a comprehensive assessment of the impact of this technological process on the amino acid composition, biological value, and in vitro digestion kinetics of the obtained peptide fractions in comparison with intact proteins. Native and heat-denatured whey protein concentrates were hydrolyzed using a two-enzyme system (Alcalase and Formea CTL) and fractionated by ultrafiltration, nanofiltration, and reverse osmosis. The key results showed that the developed cascade-selective ultra-nanodiafiltration process effectively removes highmolecular-weight protein compounds, minerals, and free amino acids. The hydrolysates retain a peptide fraction with a molecular weight of 0.4–5 kDa, which exhibits maximal antioxidant (1280 and 979 µM TE/g protein equivalent) and ACE-inhibitory (IC50 =0.10 and 0.19 mg/mL) activities. The purified hydrolysates were characterized by a balanced amino acid profile (EAA/NEAA ratio of 0.77–0.78), a high content of BCAA (19.07–19.20 g/100 g protein), and an increased coefficient of utility (0.71–0.72) compared to the initial whey protein concentrates. Digestibility assessment using the INFOGEST 2.0 model confirmed high bioavailability, as well as faster and more complete digestion of the fractionated hydrolysates compared to intact proteins. The obtained results open prospects for the industrial scaling of the technology for producing whey protein hydrolysates and their use as functional ingredients. Such ingredients can be applied in the development of functional and specialized food products, including protein formulas for sports nutrition, nutritional support for metabolic syndrome and age-related sarcopenia, as well as for the correction of chronic oxidative stress and the prevention of hypertension.

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