Nowadays it is still of great demand to find a feasible, inexpensive and green material for industrial enzyme immobilization for wastewater treatment. In this work, horseradish peroxidase (HRP) was successfully immobilized on glutaraldehyde activated carbon nanospheres. Compared with free HRP, the immobilized one showed better tolerance of temperature and pH. Meanwhile the storability and reusability were also greatly improved after immobilization. The immobilized HRP could be used for removal of several phenolic compounds from an aqueous solution and exhibited excellent removal efficiency, especially for chlorophenols, 4‐methoxyphenol, and bisphenol A. The immobilization could also make the enzyme more tolerant of ions and preserve its bioactivity. Moreover, by the introduction of support‐enzyme interaction a remarkably enhanced biodegradation was observed. The biodegradation of phenols with electron‐donor groups were found to be doubled after immobilization on active carbon nanospheres. These findings might be useful for further construction of immobilized enzymes for industrial applications, which exhibit excellent stability together with high activity.