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Therefore, this work focused on fine-tuning the heterotrophic medium composition to cultivate novel green, yellow, and white <jats:italic>Chlorella vulgaris<\/jats:italic> mutant strains. Screening assays were carried out to select the most significant factors, and different nutrient concentrations were optimized by modelling biomass and protein productivity, specific growth rate, and color, via response surface methodology. The biomass and protein productivities achieved by these strains were improved by up to 70% and 94%, respectively. Additionally, biomass color was correlated with medium composition for the first time, allowing the improvement of the yellow and white mutant colorations by 20%. 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