The purpose of this study was to determine the ability of the nonbasidiomycete, filamentous fungi Penicillium frequentans, isolated and grown on sugar cane bagasse pith, to remove phenanthrene in a solidstate culture. Additionally, the study investigated whether phenanthrene removal could be enhanced by manipulating the carbon-to-nitrogen ratio (C:N) and moisture content over a relatively short-term period (29 days). To evaluate the combined effect of moisture content and the C:N ratio, a combined experimental design, composed with a 2 2 factorial and both central and axial points, was used. It was shown that the moisture content (p < 0.0003) and the combined effect of moisture content and C:N ratio have a significant (p < 0.002) positive effect on the phenanthrene removal. It was also found that heterotrophic activity was not correlated to phenanthrene removal. An optimum phenanthrene removal efficiency of 74% was realized at a moisture content of 40% and a C:N ratio of 60. This suggests that Penicillium frequentans was able to effectively remove phenanthrene in a solidstate culture and that the combination of nutrient addition and moisture adjustment could enhance the phenanthrene removal activity.