Anion carboxymethylated β-glucan alleviates undesirable binding between procyanidins and β-galactosidase.

To solve the potential problem of hindered β-galactosidase activity by procyanidins, carboxymethylated Pachyman (CMP), a negatively-charged carboxymethylated (1 → 3)-β-d-glucan, was applied to mitigate inhibition by procyanidins. The mechanisms underlying this effect were explored through enzyme kinetic analysis, fluorescence quenching assays, circular dichroism, and molecular docking studies. The results indicated that the introduction of CMP could decrease the inhibition rate of high-concentration lotus seedpod oligomeric procyanidins (LSOPC) from 98.7 to 46.5%, and enabled low-concentration LSOPC to activate β-galactosidase in vitro and in vivo. The competitive/noncompetitive inhibition constants, fluorescence quenching constants, and molecular docking results indicated that the mechanism of this effect might be CMP competing with β-galactosidase to bind procyanidins, resulting in restoration of the catalytic centre and key active site of procyanidin-bound lactase. Additionally, it was affected by procyanidin-CMP noncovalent interactions. This study illustrates a promising strategy for mitigating the anti-nutritional properties of procyanidins and activating β-galactosidase to promote intestinal health.