Valorization of Paneer waste Whey through Fermentation with Pediococcus pentosaceus NCDC 273 Insights from Intracellular Metabolomics by GC-MS

The present study employed an untargeted GC-MS-based metabolomics approach to investigate the intracellular metabolic landscape of Pediococcus pentosaceus NCDC 273 during the fermentation of paneer whey. The goal was to understand the metabolic dynamics that enable this bacterium to efficiently utilize a dairy by-product and produce value-added compounds. Intracellular metabolites were profiled at the early (6 h) and late (18 h) exponential growth phases, revealing a distinct temporal metabolic reprogramming. Multivariate analysis and correlation networks confirmed significant metabolic shifts, where the early phase was characterized by active uptake and utilization of diverse sugars and fatty acids to support rapid proliferation. In contrast, the late phase demonstrated a metabolic reorientation towards energy production, redox balance, and stress adaptation. This was evidenced by the significant accumulation of key metabolites such as lactic acid, nicotinamide, and trehalose, which are crucial for maintaining growth and tolerance in an acidifying environment. Overall, the findings demonstrate that P. pentosaceus NCDC 273 possesses the metabolic flexibility to effectively channel whey-derived sugars into central metabolism, while concurrently deploying adaptive strategies for survival. This research represents the first intracellular metabolomic characterization of this strain during whey fermentation, providing novel mechanistic insights that reinforce its potential for the valorization of paneer whey into functional bioproducts.