The highly porous Mn2O3-Ag nanofibers were fabricated by a facile two-step procedure (electrospinning and calcination). The structure and composition of the Mn2O3-Ag nanofibers were characterized by SEM, TEM, XRD, EDX and SAED. The as-prepared Mn2O3-Ag nanofibers were then employed as the immobilization matrix for glucose oxidase (GOD) to construct an amperometric glucose biosensor. The biosensor shows fast response to glucose, high sensitivity (40.60 mu A mM(-1) cm(-2)), low detection limit (1.73 mu M at S/N=3), low K-m.app value and excellent selectivity. These results indicate that the novel Mn2O3-Ag nanfibers-GOD composite has great potential application in oxygen-reduction based glucose biosensing.