OPTIMIZATION OF PREBIOTIC RATIO FOR IMPROVED BSH ACTIVITY OF ENTEROCOCCUS FAECALIS CGZ3

Abstract

This study aimed to optimize prebiotic formulations to enhance bile salt hydrolase (BSH) activity in Enterococcus faecalis CGz3, a promising probiotic strain, using a one-factor-at-a-time (OFAT) approach followed by response surface methodology (RSM). Prebiotics such as Taraxacum officinale (dandelion) root extract, inulin, fructooligosaccharides, sorbitol, and maltitol were tested for their effects on BSH activity. A Box-Behnken design identified an optimal blend of sorbitol (0.4 mg/mL, v/v), maltitol (0.2 mg/mL), and dandelion root extract (0.5 mg/mL), achieving a BSH activity of 182.5 mg/mL. Dandelion root extract was prepared via Soxhlet extraction at 80°C using polar and non-polar solvents and analyzed for total phenolic content (TPC), total carotenoid content (TCC), and antioxidant capacity via the DPPH assay. Gas chromatography-mass spectrometry (GC-MS) detected 25 bioactive compounds, and high-performance liquid chromatography (HPLC) quantified chlorogenic acid, a potent polyphenol. These findings highlight the potential of plant-based prebiotics in synbiotic development and provide a novel approach for optimizing prebiotic ratios to support cholesterol metabolism through BSH activity.