Aluminum pillared interlayered clay (Al-PILC) calcined at 773 K was the most appropriate support for immobilization of horseradish peroxidase (HRP) among several materials investigated.HRP could be immobilized on the Al-PILC via hydrogen bonding between functional groups of support and amino groups of the enzyme.The support and immobilized enzyme were characterized with N2-adsorption and desorption, XRD and FT-IR.An optimum condition for immobilization with enzyme immobilization efficiency of about 100% was obtained when the dose of support was greater than 0.45 mg per unit enzyme activity and reaction time exceeded 15 min.HRP immobilized on Al-PILC exhibited good catalytic performance.