Nanoporous silica nanofibers have been employed as a matrix to encapsulate horseradish peroxide enzymes via a simple electrospinning method. A viscous solution of prehydrolyzed tetramethyl orthosilicate, beta-d-glucose, poly(vinyl alcohol), and enzymes were employed as spinning solution to generate porous fibers in the form of nonwoven mats. The silica fiber mats thus produced have a high surface area because of the small diameter (100 to 200 nm) of the fibers as well as the extreme porosity (2 to 4 nm) of individual fibers caused by the glucose template present in them. The high surface area, mechanical flexibility, thermal stability, reusability, and freedom of encapsulating various enzymes make porous silica nanofibers excellent biosensors.