| Summary: | This study investigates the impact of urea and β-GP on the growth of <i>Streptococcus thermophilus</i> S-3, a bacterium commonly used in industrial fermentation processes. Through a series of growth experiments, transcriptome, metabolome, and omics-based analyses, the research demonstrates that both urea and β-GP can enhance the biomass of <i>S. thermophilus</i>, with urea showing a more significant effect. The optimal urea concentration for growth was determined to be 3 g/L in M17 medium. The study also highlights the metabolic pathways influenced by urea and β-GP, particularly the galactose metabolism pathway, which is crucial for cell growth when lactose is the substrate. The integration of omics data into the genome-scale metabolic model of <i>S. thermophilus</i>, iCH502, allowed for a more accurate prediction of metabolic fluxes and growth rates. The study concludes that urea can serve as a viable substitute for β-GP in the cultivation of <i>S. thermophilus</i>, offering potential cost and efficiency benefits in industrial fermentation processes. The findings are supported by validation experiments with 11 additional strains of <i>S. thermophilus</i>, which showed increased biomass in UM17 medium.
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