Glycerol is an attractive fuel for a fuel cell, because it is non-toxic, non-volatile, non-flammable, has high energy density, and is abundant due to the fact that it is a byproduct of biodiesel production. However, it has not been an effective fuel for low temperature, precious metal catalyzed fuel cells. In this paper, we describe the use of glycerol as a fuel in an enzymatic biofuel cell. An alcohol dehydrogenase and aldehyde dehydrogenase-based bioanode has been developed that oxidizes glycerol, a safe high energy density fuel. Glycerol/O-2 biofuel cells employing these bioanodes have yielded power densities of up to 1.21 mW cm(-2), and have the ability to operate at 98.9% fuel concentrations. Previous biofuel cells could not operate effectively at high fuel concentrations due to the nature of the solid fuel such as sugar or the solvent characteristics of fuels such as lower aliphatic alcohols. The glycerol/O-2 biofuel cell provides improved power densities compared to ethanol biofuel cells due to ability to more completely oxidize the fuel. (C) 2007 Elsevier B.V. All rights reserved.