<p>Tofu industries have developed quite rapidly in small and medium scale industries with a total of around 86,400 units producing of more than 2.56&#xa0;million tons per year. The increasing number of tofu industries render problems in the environment since it contributes up to 20&#xa0;million m³ of liquid waste per year. The aim of this research was to utilize TW as the medium for <i>Botryococcus braunii</i> on mixotrophic condition to produce lipid. The effluent was varied at different concentrations from 5 to 20% (v/v). The growth rate, biomass productivity, total lipid and reduced organic levels of COD were also investigated at varying glucose and glycerol concentrations (g L^− 1) with intervals from 15:0 to 3:16&#xa0;g L^− 1. Results showed that the optimum growth of <i>Botryococcus braunii</i> was found at 10% (v/v) of TW and 12:4 (glucose: glycerol), which resulted for 6.57 ± 0.03&#xa0;g L^− 1 biomass, 1.48 ± 0.02 day^− 1 growth rate and 90% of COD reduction efficiency (<i>p</i> &lt; 0.05). Whereas, the highest lipid content was achieved at 6:12&#xa0;g L^− 1 (glucose: glycerol), 41.13 ± 0.74% (<i>p</i> &lt; 0.05). These findings demonstrate that TW has the capacity to yield valuable microalgal biomass and lipid, which can be used to produce lucrative products.</p> Graphical Abstract <p></p>