An alkane-biodegrading bacterium identified as Rhodococcus erythropolis (NTU-1 strain) was isolated from petroleum contaminated soil. The major purpose of the current research was to study the issues regarding biofloccules formation and cell surface hydrophobicity of NTU-1. When long-chain alkanes are supplied as the carbon source, NTU-1 tends to form biofloccules and remove significant amount of alkanes by biodegradation and physical trapping. Approximately, more than 95% of each alkane could be efficiently removed within 40-68 h. The bioremediation process was accompanied by formation of biofloccules with size ranging from 0.1 to 2 cm in diameter. The MATH test and the hydrophobic slide experiment suggested that NTU-1 might possess a hydrophobic cell surface which is one of the important factors in the formation of biofloccules. It provides the interaction of cells with hydrocarbon droplets effectively and further aggregate into larger clumps. Besides, when grown on n-hexadecane, experimental results revealed that there were at least 11 different growth-associated fatty acids produced, with carbon chain length ranging from 12 to 24, and cell surface hydrophobicity was enhanced via accumulation at the cell surface. (C) 2009 Elsevier Ltd. All rights reserved.