In this study the lipid production potential of the isolated yeast Rhodotorula glutinis TR29 in molasses medium under non-aseptic culture conditions was investigated. Different molasses concentrations and initial pH values were tested to make R. glutinis TR29 cells more dominant population in the medium, thereby preventing undesired microbial contaminants. Contamination could be prevented by selecting the high molasses concentration (20%) and low initial pH (5.0). When these parameters were kept constant, the optimum temperature, additional nitrogen source concentration and incubation time for the lipid production were found to be 25 degrees C, 4 g/L and 168 h, respectively. Under these culture conditions, the cell mass and lipid concentration were determined as 16.2 and 10.5 g/L, respectively. The lipid content was determined as 64.8%. The main cellular fatty acids of the yeast were oleic (63.5%), politic acid (15.4%), stearic acid (9.1%) and palmiteloic acid (7.2%). The yeast lipids seems to be a promising feedstock for biodiesel production due to a high content of C16 and C18 fatty acids. To our best knowledge, this is the first work on lipid production by yeasts in non-sterile molasses medium. (C) 2016 Elsevier Ltd. All rights reserved.