Recent research has highlighted the effectiveness of starter inocula in fermentation processes. In this sense, this study examines the use of an inoculum composed of coffee pulp, mucilage broth, and microorganisms such as Saccharomyces cerevisiae, L. delbrueckii ssp. bulgaricus, and S. thermophilus in fermenting Castillo variety coffee. An inoculum was prepared, measuring variables such as the pH, acidity, °Brix, lactic acid bacteria, and yeast viability. Following optimization, the inoculum was evaluated in a fermentation process, evaluating the pH, °Brix, acidity, microbiological analysis, ochratoxin A, and cup quality post-drying and roasting. The findings demonstrated a significant reduction in the pH from 4.47 to 4.05 and in the °Brix from 15.8 to 8.45, indicating efficient organic acid production and sugar degradation. Acidity levels increased from 20.02 mg/g to 42.69 mg/g, while microbial viabilities remained above 107 CFUs/g, suggesting effective biomass production. The process effectively reduced the microbial load without detecting ochratoxin A. Sensory evaluations confirmed the enhanced cup quality, validating the positive impact of inoculum use in coffee fermentation. The results support the use of coffee pulp and mucilage broth as effective substrates for the growth of the evaluated microorganisms, and the application of starter cultures containing lactic acid bacteria and yeast can elevate the coffee to a specialty grade.