The aim of this study was to synthesize, characterize, and observe the catalytic activity of Pd1Au1 supported by vapor-grown carbon nanofiber (VGCNF) anode catalyst prepared via the chemical reduction method. The formation of the single-phase compounds was confirmed by X-ray diffraction (XRD) and Rietveld refinement analysis, which showed single peaks corresponding to the (111) plane of the cubic crystal structure. Further analysis was carried out by field emission scanning emission microscopy (FESEM), energy dispersive X-ray analysis (EDX), nitrogen adsorption/desorption measurements, and X-ray photoelectron spectroscopy (XPS). The electrochemical performance was examined by cyclic voltammetry tests. The presence of mesoporous VGCNF as support enables the use of a relatively small amount of metal catalyst that still produces an excellent current density (66.33 mA cm-2). Furthermore, the assessment of the kinetic activity of the nanocatalyst using the Tafel plot suggests that Pd1Au1/VGCNF exerts a strong electrocatalytic effect in glycerol oxidation reactions. The engineering challenges are apparent from the fact that the application of the homemade anode catalyst to the passive direct glycerol fuel cell shows the power density of only 3.9 mW cm-2. To understand the low performance, FESEM observation of the membrane electrode assembly (MEA) was carried out, examining several morphological defects that play a crucial role and affect the performance of the direct glycerol fuel cell.