Improvement of lactose digestion by highland barley (Hordeum vulgare var. coeleste L.) β-glucan: Activation of lactase under simulated gastric/small intestinal digestive conditions

β-Galactosidase (lactase) plays a crucial role as a dietary supplement in managing lactose intolerance. Here, the catalytic activity of lactase was successfully activated for the first time through complexation with water-extractable β-glucans from highland barley (WHBG). Under simulated gastric/small intestinal digestive conditions, WHBG and lactase spontaneously formed complexes, resulting in a remarkable increase in catalytic activity up to 172.6 %. Structural analyses revealed that the incorporation of WHBG caused partial unfolding of lactase, thereby exposing its hydrophobic regions with active sites, and the electrostatic and hydrophobic interactions between the two played pivotal roles. Meanwhile, according to microstructure and particle size analyses, the dissociation of aggregates and the re-distribution of lactase molecules were also observed. Consequently, the enzyme-substrate contact was promoted, and the hydrolysis efficiency of complexed lactase in the digestion of lactose in milk was superior to that of native lactase. Notably, among WHBG30/50/70 obtained by continuous fractionation of WHBG with 30 %/50 %/70 % ethanol, WHBG70 exhibited the lowest molecular weights and size, and the highest negative ζ-potential, potentially contributing to its superior activation abilities on lactase. These findings challenge the traditional view of polysaccharides as enzyme inhibitors and highlight their potential for diverse applications.