Abstract In this research, statistical optimization designs were applied to evaluate the growth of the Chlorella vulgaris in mixotrophic cultures enriched with nitrogen and phosphorus. The influence of each factor was evaluated at three levels, so a multifactorial experimental design was created 32. The results obtained showed that microalgae growth improves 32% by adding 50 mg of nitrogen and 25 mg of Nitrogen to the culture. This project demonstrates the relevance of the design of experiments for the optimization of algal biomass production in order to obtain bioproducts.