The application of a crude enzymatic extract from the mixed culture of Trametes maxima (T. maxima) and Paecilomyces carneus (P. carneus) in removal of phenanthrene (PHE) from water was assessed. Production of crude extract from a mixed culture of T. maxima and P. carneus does not require special conditions yet the use of 20 g/L of glycerol as carbon source enhances laccase and manganese-dependent peroxidase activities. Application of crude extracts became feasible since it does not require further purification. The crude enzymes entrapped in alginate beads in a fixed-bed packed column reached 94.3%-0.7% of PHE removal after 13 hr of continuous treatment under nonsterile conditions, while the alginate beads without enzymes reached only 45.3%-23% of PHE removal. The difference between the removal of alginate beads with and without enzymes was the oxidative activity on the surface of the alginate beads, which was observed in the increase in the C=O group and in the X-ray spectroscopy analyses of the C 1s and O 1s regions. Beads without enzymes had less C=O groups. Polyoxyethylene sorbitan monooleate and dimethyl sulfoxide influenced enzymatic activity to carry out a level of PHE removal similar to a photocatalytic treatment.