Fungal Pectinolytic Enzyme System for the Production of Long- and Short-Chain Pectin-Derived Oligosaccharides (POS) from Pomelo Albedo and Their Prebiotic Potential

Pectin-derived oligosaccharides (POS) are emerging as promising functional prebiotics with growing industrial interest. This study reports a synergistic fungal pectinolytic biocatalytic system comprising endopolygalacturonase (EndoPG) and pectin methylesterase (PET11) from Aspergillus aculeatinus BCC 17849 for the controlled depolymerization of pomelo (Citrus maxima) albedo pectin. PET11-mediated demethylation increased substrate accessibility, thereby enhancing EndoPG-catalyzed hydrolysis and resulting in higher POS yields than those obtained with single-enzyme systems. The highest production of short-chain POS, comprising GalA, di-GalA, and tri-GalA (681 mg/g substrate), was achieved at an EndoPG:PET11 dosage ratio of 15:5. The resulting POS fraction significantly promoted the growth of five probiotic strains, including Lactobacilli and Bifidobacteria species, and enhanced probiotic adherence to intestinal epithelial cells. In particular, Lactobacillus acidophilus TBRC 5030 exhibited the highest adhesion level (35.24 ± 6.43%) in the presence of 2.0 mg/mL POS. Overall, this work demonstrated that enzyme-assisted demethylation coupled with targeted endo-hydrolysis enables effective tailoring of POS chain length, providing a promising biocatalytic strategy for pectin valorization into prebiotic ingredients.