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Vinasse is ethanol wastewater that still contains nutrient so that it can be medium cultivation for chlorella sp. Cultivation chlorella sp. in vinasse not only for minimize its COD content but potentially to produce PHB in limited nutrient. This paper presents a cultivation process of chlorella sp in vinasse on various limited nutrient. Chlorella sp was cultivated in vinasse by adding complete nutrient (urea and TSP); adding TSP in vinasse (limited N) and adding urea in vinasse (limited P). Experiment data was optimized by mathematical model to compare the difference of limited nutrient systematically. The study confirmed that the best condition of medium of microalgae cultivation to minimize COD content in vinasse by addition phosphate into reactor. However, PHB could be best accumulated in the cell on limited phosphate medium.