Textile discharge printing is the most versatile and important of the methods used for introducing design to textile fabrics. In discharge styles, the pattern is produced by the chemical destruction of the original dye in the printed areas. The discharging agents used can be oxidising or reducing agents, acids, alkalis and various salts. However, the most important methods of discharging are based formaldehyde sulphoxylates and thiourea dioxide. Recently, environmental and industrial safety concerns have increased the potential for the use of enzymes in textile processing to ensure eco-friendly production. Formaldehyde sulphoxylate (NaHSO2.CH2O.2H2O) is one of the most powerful discharging agents; however, it is quite toxic and produces formaldehyde, a known human carcinogen associated with nasal sinus cancer and nasopharyngeal cancer. In this work, a hazardous chemical has been replaced with eco-friendly horseradish peroxidase enzyme in textile discharge printing. Enzymatic discharge printing was carried out with a phenol oxidising enzyme system such that the reactive dye was selectively discharged from the cotton fabric in selected areas, creating a printed surface. The effects of enzyme concentration, pH of the printing paste, treatment time and the temperature of enzymatic treatment were studied. The optimum conditions for enzymatic discharge printing were found to be pH 8.5 at 70°C with a dye concentration of 80 g/L and 60 min as the treatment time.