A single-stage mixotrophic cultivation strategy was developed with a novel microalgal isolate Chlorella sorokiniana FC6 IITG for high cell density lipid-rich biomass generation. The strain was evaluated for growth and lipid content under different physico-chemical parameters, nutritional conditions and trophic modes. Finally, a single-stage mixotrophic fed-batch cultivation strategy was demonstrated with intermittent feeding of key nutrients along with dynamic increase in light intensity for high cell density biomass and sodium acetate as elicitor for lipid enrichment. The key findings: (i) glucose and sodium acetate was identified as growth supporting and lipid inducing nutrients, respectively; (ii) mixotrophic batch cultivation resulted in maximum biomass and lipid productivity (mg L-1 day(-1)) of 455.5 and 111.85, respectively; (iii) single-stage mixotrophic fed-batch cultivation showed maximum biomass productivity of 1.93 g L-1 day(-1) (biomass titer 15.81 g L-1) and lipid productivity of 550 mg L-1 day(-1); (iv) biodiesel properties were in accordance with international standards. (C) 2014 Elsevier Ltd. All rights reserved.