Coupling dark fermentation (DF), which produces hydrogen from diverse effluents or solid waste, and heterotrophic cultivation of microalgae, which produces lipids, carbohydrates and proteins, is a promising and innovative solution for developing sustainable biorefineries. The use of a raw DF effluent, containing acetate and butyrate, to support the heterotrophic growth of Chlorella sorokiniana was investigated. All the acetate in sterilized and unsterilized DF effluent was exhausted in less than three days of heterotrophic cultivation, whereas butyrate was not used by the microalgae. The microalgae biomass reached 0.33 g L− 1 with a carbon yield on acetate of 55%. The algal yield was higher than previously reported for synthetic DF effluent. It was concluded that compounds other than volatile fatty acids were present in the DF effluent and these could be consumed by the microalgae. After the acetate had been exhausted, butyrate was consumed by facultative and strict aerobic bacteria originating from the DF effluent. The concentration of the bacterial community increased during the experiment but did not have any significant impact on heterotrophic microalgae growth. A high microalgal biomass yield was achieved without requiring the DF effluent to be sterilized.