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Abstract: Carboxyl-modified mesoporous materials have been demonstrated as efficient carriers for immobilizing enzymes. However, little attention has been paid to the effect of H2SO4 treatment, a key in the synthesis of carboxyl-modified carriers, on the immobilization. To examine the H2SO4 treatment’s influence, we contrasted the ability of immobilizing papain between SBA-15 and the one after H2SO4 treatment (H2SO4-treated SBA-15). Meanwhile, the contribution of the as-synthesized carboxyl to the immobilization was also rechecked by comparing carboxyl-modified SBA-15 (COOH-SBA-15) and H2SO4-treated SBA-15. In relation to the experiments, the H2SO4 treatment increases the immobilizing ability of SBA-15, especially when the immobilization performs near the pI of papain. Moreover, the as-synthesized carboxyl facilitates the immobilization by means of strengthening the hydrophobic dehydration at the interface, instead of merely through electrostatic forces. Additionally, the carboxyl makes a large contribution to inducing the favorable conformation change of papain, improving the specific activity, catalytic efficiency, and binding affinity of the immobilized enzyme when combined with the substrate. [Copyright &y& Elsevier]