This study evaluates the potential of the enzyme horseradish peroxidase in the decolorization of one common industrial azo dye, naphthol blue black. Studies are carried out to understand the process parameters such as pH, temperature and reaction time. The enzymatic decolorization of the dye was examined by UV-Vis spectrophotometer and LC-MS measurements. Temperature and pH conditions were optimized for obtaining high azo-dye decolorization. Azo-dye removal at a pH range 4-6 was found to be the highest for all temperatures. After 5 minutes of treatment, the color removal of dye was ca. 80-90%. The LC-MS and spectrophotometric analyses indicated that the decolorization of the azo dye with enzyme was due to the reduction of the azo bonds. This study verifies the viability of the use of the horseradish peroxidase in the decolorization of naphthol blue black.