Degradation of phenolics by members of soil microflora is an important means by which these substances are removed from the environment thus reducing environmental pollution. Biodegradation by microorganisms offers unique opportunities to destroy or render phenolic compounds. A bacterium, PU1, identified as Pseudomonas fluorescens PU1, was investigated for its ability to grow on and degrade phenols as sole carbon sources in aerobic shaking batch culture. The organism degraded up to 1000 ppm of phenol using meta cleavage pathway. The pathways for phenol degradation were proposed by the identification of metabolites and assay of ring cleavage enzymes in cell extracts. Phenol was degraded via catechol with subsequent metaring cleavage. Cell growth increased as the phenol concentrations increased up to 1000 ppm phenol. The biodegradation efficiency, degradation extent, and metabolic pathway of phenol were determined to provide useful clues for further application of this isolate in the engineered bioremediation systems. The paper's results suggest that Pseudomonas fluorescens PU1 strain could be a good candidate for remediation of phenol contaminants from heavily polluted sites.