The study aimed to optimize the process parameters for the production of virgin coconut oil (VCO) by exploring the use of aerobic fermented neera (coconut inflorescence sap) at different concentrations (ranging from 5% to 25%). Fermented neera can be prepared from fresh neera without incurring additional expenses, presenting a potential advantage over commercially available cultures in the market. Coconut milk was extracted from a coconut kernel using a hydraulic coconut milk expeller. The extracted coconut milk was then fermented with varied concentrations of fermented neera ranging from 5% to 25%, and then, the oil layer was separated. Fermented neera as a starter at a concentration of 10% resulted in the highest VCO yield of 19.8% following 14 hours of fermentation. The study also found that the fermentation duration of the starter (6-hour, 8-hour, and 10-hour fermented neera) and its concentration had significant effects on the yield of VCO. However, the effects of starter on the physical properties (moisture content, viscosity, specific gravity, and refractive index) and biochemical profile (free fatty acid, peroxide value, acid value, saponification value, iodine value, and total phenolic content) of the resultant VCO were not significant. The physicochemical properties of the VCO produced using the fermented neera-based coconut milk fermentation method met the standards stipulated by the Food Safety and Standards Authority of India (FSSAI) and the Asian and Pacific Coconut Community (APCC). Thus, fermented neera offers a novel economical alternative to enhance the fermentation rate during VCO production.