Graphene oxide (GO) has shown promising applications as supports for heterogeneous catalysts. This study reports the functionalization of GO by (3-mercaptopropyl) trimethoxysilane to immobilize lipase. The immobilization is carried out by the site-specific covalent binding between thiol groups of lipase and thiolsulfonate groups on modified GO. For the hydrolysis of p-nitrophenylpalmitate, the obtained GOlipase shows catalytic performance (93.4%) comparable with that of the free lipase, and the reusability after 10 times is 69.9%. Compared with the free lipase, the GOlipase exhibits good pH, thermal, and storage stability, which are important in practical applications.