Enzyme Treated Sapota Seed Dietary Fibre Is a Novel Prebiotic Feed Additive: In Vitro Studies

The valorization of agro-industrial fruit by-products presents a sustainable strategy to enhance animal nutrition while reducing environmental waste. This study investigates the physicochemical attributes, dietary fiber profile, and prebiotic potential of the enzyme treated Manilkara zapota (sapota) seed powder (eSSP) for functional use in poultry feed. The eSSP flour demonstrated high crude fiber content (23.94 ± 1.86 g/100 g), with total dietary fiber comprising 83.45% insoluble and 16.54% soluble fractions. Enzymatic hydrolysis optimized at 6 h revealed peak concentrations of fermentable oligosaccharides, including galacto-oligosaccharides (12.06 ± 0.45%), manno-oligosaccharides (8.04 ± 0.30%), fructo-oligosaccharides (9.83 ± 0.25%), and xylo-oligosaccharides (10.83 ± 0.50%). Supplementation with e₆SSP resulted in a significant increase in both qualitative and quantitative volatile fatty acid (VFA) production, indicating its prebiotic potential. Notably, the high xylo-oligosaccharide (XOS) content (∼10%) contributed to elevated butyric acid levels in fermentation assays, reinforcing the stimbiotic properties of eSSP. Symbiotic assays with Lactobacillus casei confirmed the eSSP's capacity to support probiotic growth, while in vitro fermentation demonstrated enhanced production of short-chain fatty acids (SCFAs), particularly butyrate. Antioxidant profiling further validated the seed's bioactive potential, with total phenolic content of 767.65 ± 1.24 mg GAE/100 g and flavonoid content of 2223.6 ± 0.87 mg QE/100 g. These findings establish eSSP as a potent, cost-effective, and natural prebiotic candidate for improving gut health and sustainability in animal feed systems.