This study presents the development and characterization of a novel buffalo Petit Suisse cheese, enhanced with symbiotic properties through an innovative whey retention method and incorporating inulin and xanthan gum. The research focused on assessing the cheese’s physicochemical properties, shelf life, lactic acid bacteria viability, syneresis behavior, and the impact of varying concentrations of functional ingredients. The addition of inulin and xanthan gum, following a design of experiments, significantly influenced the cheese’s texture and consistency. Higher inulin concentrations were associated with increased fermentation activity, as indicated by total titratable acidity, which showed an increase from 1.22% to 1.50% over a 28-day period, and pH levels that decreased from 3.33 to 2.96. The syneresis index varied across trials, with the highest reduction observed in trials with increased xanthan gum concentrations, effectively reducing syneresis to 0%. Lactic acid bacteria viability also showed notable variations, with the highest cell survival percentage reaching 107.89% in formulations with higher inulin and xanthan gum concentrations. These results underscore the importance of inulin and xanthan gum in enhancing the cheese’s microbial stability and textural quality. The study concludes that the strategic use of inulin and xanthan gum improves the nutritional profile of buffalo Petit Suisse cheese and optimizes its textural and sensory attributes.