A fluorene-degrading strain named USTB-Cwas isolated from crudeoil of production well inDagang oil field, southeast of Tianjin, northeast China. The 16S rDNA sequence of USTB-Cshowed 100% similarity with that of RhodococcuserythropolisPR4, it was belonged to Rhodococcus. The strain USTB-Ccould remove 55.8% of fluorenewith an initial concentration of 100 mg L-1in 23days without other substrates.It implied that the strain USTB-C showed the potential to degrading efficient of fluorene.Based on the intermediatemetabolitesanalyzed by gas chromatography-mass spectrometry(GC-MS), we deduced the possible metabolic pathway of strain USTB-Cfor fluorenebiodegradation. Fluorene was initially oxidized at C-9positions finally resulting 9-fluorenone. Through hydrolysisreaction, 9-fluorene subject to form phthalic acid. Furthermore, phthalic acid was further degraded to1 -hydroxybenzene. USTB-C evidently possessesefficient, high effective degrader and potential for further application on the enhanced bioremediationtechnologies for treating fluorene-contaminated soil.