Green algae produce fatty acid components that can be used in biofuel production without the need for additional nutrients. We aimed to elucidate the contribution of the T. obliquus KNUA019 through the control of the optimal culture (solvent, culture time, nitrogen, and phosphorus) conditions and thermal analysis (DTA and TGA curves) that affects fatty acid productivity maximum lipid yields from the four culture methods. Phylogenetic analysis of Tetradesmus obliquus (Turpin), Scenedesmus obliquus (Turpin), Acutodesmus obliquus (Turpin), and Chlorella sorokiniana (Shihira & R.W.Krauss) strains was attempted using the internal transcribed spacer. T. obliquus KNUA019 can produce significant amounts of carbon-containing components, which are valuable for use as energy sources. As a result of GC analysis, T. obliquus KNUA019 generates fatty acid components that are directly useful as biofuels, such as tetradecanoic acid (C 14 H 28 O 2), methyl Z-11-tetradecenoate (C 15 H 28 O 2), tetradecanoic acid (C 15 H 30 O 2), 9-hexadecenoic acid (C 15 H 30 O 2), pentadecane (C 15 H 32), 8-heptadecene (C 17 H 34), hexadecanoic acid (C 17 H 34 O 2), heptadecane (C 17 H 36), 9-octadecenoic acid (C 19 H 36 O 2), octadecanoic acid (C 19 H 38 O 2), and 3,7,11,15-tetramethyl-2-hexadecen-1-ol (C 20 H 40 O). These fatty acids can be used directly as biofuel precursors without transesterification. We indicate that commercial biofuel production is possible using mass culture of T. obliquus KNUA019, reducing production costs. This process was indicated as an optimum method for simplifying the process of fatty acid components under optimal culture conditions for a source of biofuels. Keywords Biomass • Fatty acid • Green algae • Tetradesmus obliquus Young-Saeng Kim and Jung Yi have contributed equally to this work.