An enzymatic nanoreactor combining the advantages of speediness, sensitivity and stability based on the macroporous ordered siliceous foam (MOSF) is proposed in this work. It was structured through the electrostatic interaction matching between the biomolecules and porous materials after 30 min incubation of MOSF in trypsin solution. Myoglobin as the model protein was used to validate the nanoreactor, and the tryptic products were analyzed by MALDI-TOF mass spectrometry. Comparing with the standard in-solution digestion results, proteolysis assisted by trypsin-MOSF nanoreactor is much more efficient due to the large surface areas, interspace confinement effect and enrichment of the substrates within the reactor pores. The nanoreactor showed satisfactory sensitivity even the concentrations of myoglobin lowed to 2 ng/μl. The subsequent tests indicates that the reactor is stable enough to retain its high enzymatic activity even refrigerated for several weeks or interfered by a certain concentration of surfactants or salts, and would bring out conveniences for many biochemical studies. A complex proteome sample was also digested by using the trypsin-MOSF reactor and then analyzed by LC-ESI mass spectrometry (LTQ, Thermal-fisher), in which 164 proteins were identified after standard protein data searching. Tripsin-immobilized MOSF nanoreactor is anticipated to greatly facilitate the characterization and understanding of the biological role of proteins.