Polyporus sp. S133, a fungus collected from contaminated-soil was used to degrade chrysene, a polycyclic aromatic hydrocarbon (PAH) in a mineral salt broth (MSB) liquid culture. Maximal degradation rate of chrysene (65%) was obtained when Polyporus sp. S133 was incubated in the cultures supplemented with polypeptone (10%) for 30 days under agitation of 120rpm, as compared to just 24% degradation rate in non-agitated culture. Furthermore, the degradation of chrysene was affected by the addition of carbon and nitrogen sources as well as kind of surfactants. The degradation rate was increased with increase in added amount of carbon and nitrogen sources, respectively. The degradation rate in agitated cultures was enhanced about 2 times higher than that in non-agitated cultures. The degradation mechanism of chrysene by Polyporus sp. S133 was determined through identification of several metabolites; chrysenequinone, 1-hydroxy-2-naphthoic acid, phthalic acid, salicylic acid, protocatechuic acid, gentisic acid, and catechol. Several enzymes (manganese peroxidase, lignin peroxidase, laccase, 1,2-dioxygenase and 2,3-dioxygenase) produced by Polyporus sp. S133 were detected during the incubation. The highest enzyme activity was shown by 1,2-dioxygenase (237.5Ul⁻¹) after 20 days of incubation.