Enhanced utilization of okara through microbial fermentation: Isolation and immunostimulatory potential of a β-galactan

Okara, a byproduct of soybean processing, can be valorized through microbial fermentation to yield bioactive compounds. In this study, we used Lacticaseibacillus paracasei , a Gram-positive lactic acid bacterium commonly applied in probiotic and dairy fermentations, to perform solid-state fermentation of okara. Crude polysaccharides were then purified using DEAE Sepharose Fast Flow and BioGel P-2 gel filtration chromatography, yielding a high-purity β -1,4-galactan polysaccharide fraction (LPP-A) with an average molecular weight of 6 kDa. To assess its immunostimulatory potential, LPP-A was evaluated as an adjuvant in a murine vaccination model with ovalbumin (OVA) as the antigen. Mice immunized with OVA plus LPP-A induced robust OVA-specific cytotoxic T lymphocyte (CTL) responses and high IgG antibody titers, indicating the induction of strong antigen-specific cellular and humoral responses. In a subsequent tumor challenge, the OVA plus LPP-A group showed significantly inhibited EG7-OVA tumor growth, suggesting high potential for LPP-A as an immune-enhancing adjuvant for cancer immunotherapy. These results demonstrate that fermenting okara can produce a β -1,4-galactan (LPP-A) with potent vaccine adjuvant properties, supporting its value in protein-based immunization strategies. Fermentation of okara by Lacticaseibacillus paracasei produced a 6 kDa β -1,4-galactan (LPP-A). As an adjuvant, LPP-A enhanced OVA-specific CTL and IgG responses, suppressed tumor growth in mice, and demonstrated potential for cancer immunotherapy.