Cell-free culture broth of Phanerochaete chrysosporium has been adopted to biologically degrade 2,4,5-trichlorophenol. Two different medium compositions of nitrogen-sufficient and nitrogen-limited were compared for their distribution of isozymes, activity of lignin peroxidase, and production of oxalate. The two different culture broths were tested for their ability to degrade 2,4,5-trichlorophenol, and the biodegradation efficiency was estimated in terms of the disappearance of 2,4,5-trichlorophenol. The degradation efficiency for the nitrogen-limited culture broth was higher than that of the nitrogen-sufficient culture broth, since the nitrogen-limited culture broth induced lignin peroxidases (LiPs) and manganese peroxidases (MnPs), and contained sufficient oxalate for producing necessary radicals. Finally, the possible mechanism of 2,4,5-CP degradation using the nitrogen-limited culture broth was proposed.