Graphene oxide (GO) nanosheets as functional material have a unique planar structure and intriguing mechanical that have attracted intensive interests recently. A method was developed for the immobilization protease on GO sheets using glutaraldehyde as cross-linking reagent. The results showed that the thermostability and reusability of immobilized protease have been obviously improved compared to the free enzyme. However, there was no significant change in optimum pH value between the free and immobilized protease. The immobilized protease exhibited good operational stability. The apparent K, and V, for free and immobilized alkaline protease were determined, and the bio-catalytic activity was not impaired by immobilization. (C) 2011 Elsevier Ltd. All rights reserved.