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The depletion of exhaustible underground petroleum resources has put the present civilization at stake, thereby warranting intense research on non-exhaustible fuel. With this energy crisis hitting the block, microorganisms such as yeasts are gaining wider importance as potential biofuel candidates. An indigenous yeast strain Saccharomyces cerevisiae isolated from laboratory-scale brewing was investigated for biodiesel production. Biodiesel was produced by in situ transesterification approach using 1,1,3,3-tetramethylguanidine as the catalyst. The fuel properties such as viscosity, density, calorific value and cetane number (CN) were determined to assess the fuel quality of S. cerevisiae biodiesel. Additionally, the investigation also focuses on theoretical studies considering the yeast de-oiled cake (low-value biomass refuse). Fatty acid methyl ester analysis revealed that biodiesel was primarily composed of tricosylic acid (C23:0, 28.71%), palmitoleic acid (C16:1, 28.96%) and oleic acid (C18:1, 18.13%). Eicosapentaenoic acid (C20:5, 2.01%), one of the most commonly known polyunsaturated fatty acid, was present in the yeast strain. The CN of yeast biodiesel was 71.58, which was much higher than petro-diesel. The theoretical findings suggest the competitiveness of yeast biomass conversion technologies with petroleum refining process economics. The overall study warrants the feasibility of co-production of biodiesel from S. cerevisiae and cracked biofuel products (from S. cerevisiae de-oiled cake) under the aegis of biorefining applications.