Currently, research is being actively conducted to utilize microalgae as biofuels due to environmental problems including global warming and energy problems due to depletion of fossil fuels. While many efforts have been made to obtain high lipid recovery from microalgae, attempts to reduce the amount of organic solvent consumed in the extraction process have been relatively few. In this thesis, we devised and optimized a new lipid extraction method that can reduce the use of organic solvents using phase separation with high lipid recovery. In addition, we carried out to utilize the remaining residue as a nitrogen source after lipid extraction. Aurantiochytrium sp. KRS101 was used as the strain and various quantitative analyzes were carried out through liquid chromatography (TLC), inductively coupled plasma spectrometer (ICP-OES), and amino acid analyzer.