Manothermonication is a recognized and efficient method used for sterilization in food preservation. The synergistic effect of sonication combined with pressure and temperature allows enhancing the cavitation activity. Never employed for extraction, this study is about the transposition of this process as a tool of extraction. In this study, Rhodosporidium toruloides yeast was submitted to extraction by four modes of sonication, with a temperature ranged from 20 to 55°C and a pressure between 1 to 2bars. The lipids extraction yields were compared to the conventional maceration. Microbial oils obtained from both processes were analyzed and quantified by HPTLC (High Performance Thin-Layer Chromatography) and GC-FID (Gas Chromatography with flame ionization detector) after transesterification of lipids. Manothermosonication (30min, 2bars, 55°C) permits to enhance of approximately 20% the extraction yield of lipids to compared to conventional maceration. The fatty acid profiles of each pretreatment and extraction by US, MS, TS and MTS do not affect the fatty acid profiles of yeast (majority of oleic acid (C18:1n9), linoleic acid (C18:2n6) and palmitic acid (C16:0)). Manothermosonication technique shows a great potential for lipid extraction from oleaginous microorganisms.