OBJECTIVE To investigate the characteristic and biochemical mechanism about the phenol biodegradation by bacterial strains ZD 4-1 and ZD 4-3. METHODS Bacterial strains ZD 4-1 and ZD 4-3 were isolated by using phenol as the sole source of carbon and energy, and identified by 16S rDNA sequence analysis. The concentrations of phenol and total organic carbon (TOC) were monitored to explore the degradation mechanism. The biodegradation intermediates were scanned at 375 nm by using a uv-vis spectrophotometer. The enzyme assays were performed to detect the activities of dioxygenases. RESULTS Bacterial strains ZD 4-1 and ZD 4-3 were identified as Comamonas testosteroni and Pseudomonas aeruginosa by 16S rDNA sequence analysis, respectively. The growth of the two strains was observed on a variety of aromatic hydrocarbons. The strains ZD 4-1 and ZD 4-3 metabolized phenol via ortho-pathways and meta-pathways, respectively. In addition, the results of enzyme assays showed that the biodegradation efficiency of phenol by meta-pathways was higher than that by ortho-pathways. Finally, the results of induction experiment indicated that the catechol dioxygenases, both catechol 1,2-dioxygenase (C120) and catechol 2,3-dioxygenase (C230), were all inducible. CONCLUSIONS The strains ZD 4-1 and ZD 4-3 metabolize phenol through ortho-pathways and meta-pathway, respectively. Furthermore, the biodegradation efficiency of phenol by meta-pathways is higher than that by ortho-pathways.