In this study, the culture medium components screening and filtering were undertaken in order to set up efficient and cost effective minimal culture media for lipid production from Yarrowia lipolytica DSM3286. The basal minimal culture medium (S2) designed yielded lipid content up to 35% of the microbial dry cell weight. A set of fermentation strategies based on this minimal medium was developed and the lipid content was raised to 51%. The scale-up under different fermentation conditions based on S2 medium led to a maximum lipid content of 65%. The produced microbial oils displayed interesting properties to be used as a feedstock for high quality biodiesel production. The minimal media and operable cultivation strategies devised in this study, in association with the works done so far by other authors, could enable fast, massive, viable and more economical production of single cell oils and smooth biodiesel manufacture.