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Algae have been explored for renewable energy, nutraceuticals, and value‐added products. However, low lipid yield is a significant impediment to its commercial viability. Genetic engineering can improve the fatty acid profile of algae without compromising its growth. This study introduced the diacylglycerol acyltransferase (BnDGAT) gene from Brassica napus into Chlorella sorokiniana‐I, a fast‐growing and thermotolerant natural strain isolated from wastewater, which increased its intracellular lipid accumulation. Hygromycin‐resistant cells were selected, and enhanced green florescence protein fluorescence was used to distinguish pure transgenic cell lines from mixed cultures. Compared to the wild type, BnDGAT expression in transgenic C. sorokiniana‐I caused a threefold increase in non‐polar lipid and a twofold increase in polyunsaturated fatty acids. Nile red staining reaffirmed the presence of higher intracellular lipid bodies in transgenic cells. There was a substantial alteration in the fatty acid profile of transgenic alga expressing BnDGAT. The non‐essential omega 9 (C18: 1) fatty acid decreased (5%–7% from 18%), while alpha‐linolenic acid, an essential omega 3 fatty acid (C18: 3), was increased (23%–24% from 11%). This study substantiates a valuable strategy for enhancing essential omega‐3 fatty acids and neutral lipids to improve its nutritional value for animal feed. The increased lipid productivity should reduce the cost of producing fatty acid methyl esters (FAME). Improved FAME quality should address the clouding issues in cold regions. [ABSTRACT FROM AUTHOR]