Contamination of soils, groundwater, air and marine environment with hazardous and toxic chemicals is major side effect by the industrialization. Bioremediation, the application of microorganism or microbial processes to degrade environmental contaminant, is one of the new environmental technologies. Because of low water solubility and volatility of diesel, bioremediation is more efficient than physical and chemical methods. The purpose of this study is biodegradation of diesel in sand by using Rhodococcus fascians, a microorganism isolated from petroleum contaminated soil. This study was performed in the column containing sand obtained from sea sides. Changes in biodegradability of diesel with various flow rates, inoculum sizes, diesel concentrations, and pH were investigated in sand column. The optimal condition for biodegradation of diesel by R. fascians in sand column system was initial pH 8 and air flow rate of 30 mL/min. Higher diesel degradation was achieved at larger inoculum size and the diesel degradation by R. fascians was not inhibited by diesel concentration up to 5%.