This study explored on evaluation of Fatty Acid Methyl Ester (FAME) profile of Chlamydomonas sp. strain TERI-1 biomass, grown mixotrophically in organic acid rich Spent Effluent (SE). Biomass productivity and FAME profile of TERI-1 strain was monitored along with its sugar and organic acid removal efficiency from SE. Maximum growth of Chlamydomonas TERI-1 strain was observed in two fold diluted (50% SE) SE. TERI-1 assimilated 10.33% sugar, 77.4% acetic acid and 6.80% butyric acid from 50% SE. FAME profile of TERI-1 biomass, cultivated in four fold diluted SE (25% SE) was composed of 21% unsaturated fatty acids and 1.8% saturated fatty acids. The unsaturated fatty acid pool was composed of 16% mono unsaturated fatty acids and 5.8% poly unsaturated fatty acid. FAME profile indicated that Chlamydomonas TERI-1 algal strain has potential for application in biodiesel production. •Chlamydomonas sp. strain TERI-1 was grown mixotrophically in organic acid rich Spent Effluent.•FAME profile of TERI-1 biomass, was composed of 21% unsaturated fatty acids.•The unsaturated fatty acid pool composed of 16% mono unsaturated fatty acids and 5.8% poly unsaturated fatty acid.•FAME profile indicated for suitability of TERI-1 strain for application in biodiesel production.