With the rapid industrialization, an ever-increasing quantity and kind of new organic compounds pose environmental problems due to their toxicity and physiological effect. However, research on the biodegradation of these compounds under anaerobic condition is very limited inspite of its efficiency and economical advantage. In this research, the pH effect on the ring cleavage of phenol under anaerobic condition was investigated, and the theory of phase separation was applied to the degradation of phenol for investigating the role of acidogenic bacteria. Results, obtained from biochemical methane potential(BMP) assay for 15.5 days of incubation, showed that acidic condition was more desirable for phenol degradation than alkaline condition. By both unacclimated methanogenic granular sludge and two mixed cultures, phenol was completely removed within six weeks of incubation with a gas conversion rate of over 86% of theoretical one. However, phenol was not degraded by unacclimated acidogenic culture, and thus it is considered as a syntrophic substrate. In case of phase separated biochemical methane potential(PSBMP) assay, in which acidogenic and methanogenic culture were seeded separately and consecutively, those that had been subjected to normal acidogens for 3 to 4 weeks showed higher gas production than those seeded with sterile or frozen culture.