High energy ultra-violet radiation was used to induce mutations in the wild type strain Chlorella sp. FC2 IITG and the mutants obtained were screened for enhanced lipid content targeted towards biodiesel production. Screening of mutants under nitrogen starved photoautotrophic condition showed five mutant strains with higher lipid content as compared to the wild type strain. Maximum neutral lipid content of 22.26% (w/w, DCW) was found for FC2-25UV mutant strain, which was 48.4% higher than that of wild type culture. Further characterization under photoautotrophic two stage high cell density cultivation of lipid rich FC2-25UV resulted in total lipid content of 68% (w/w, DCW), which was 21.43% higher than the wild type strain with a marginal improvement of 11% in the total lipid productivity. Comparison of enzyme activity assays under nitrogen starved conditions in both the strains revealed an 18.2% and 31.25% increase in acetyl CoA carboxylase and di-acyl-glycerol transferase activity of mutant. The FAME composition analysis showed 41.5% and 24% increase in the fractions of C18:2 and C18:1 in the mutant strain when compared to the wild type strain with no significant difference in the other FAME fractions. Thus, the mutant strain could be a potential candidate for biodiesel production.