The aim of the study was to examine how temperature and the pH influence the progress and efficiency of an aerobic biodegradation process, where major organic pollutants are removed from beet molasses vinasse by a mixed culture of Bacillus bacteria. It was conducted in an aerated bioreactor with a stirring system in four experimental series, each composed of five processes run at temperatures of 27, 36, 45, 54 and 63 °C. In the first and second series, medium pH was not controlled, the initial pH amounted to 6.5 and 8.0, respectively. In the third and fourth series, medium pH was controlled at 6.5 and 8.0, respectively. Under optimal conditions, the pollution load of the vinasse stillage expressed as soluble chemical oxygen demand was removed with an 88.73% efficiency. The bacterial culture assimilated all organic pollutants simultaneously, but the rate of assimilation was different. An exception was the process of betaine assimilation, which intensified only when readily available carbon sources were depleted in the medium. Synthesis and assimilation of organic acids were observed in all experiments. Advantages of the proposed method include: possibility of its use at high temperatures, and no necessity for medium pH adjustment during the process.