Traditional strategies to develop Au metal nanoparticle catalysts for glucose oxidation comprise Au nanoparticles (NPs) supported on electrode surfaces. In this work, the fabrication of a new material capable of acting as an abiotic anode for the electrooxidation of glucose was developed. The material is composed of electrospun carbon fibers containing gold nanoparticles formed in situ from a polyacrylonitrile (PAN) solution with HAuCl4. This approach allows the pre‐reduction of the gold salt by PAN under mild conditions without the need for extra energy and leads to very stable carbon–gold bonding with well‐dispersed AuNPs, not previously reported. The gold‐modified carbon fibers (Au@CFs) were characterized by SEM, energy‐dispersive X‐ray spectroscopy, XRD, and electrochemical analysis. The Au@CF electrodes showed electrochemical activity toward glucose oxidation in alkaline media. Combined to a bilirubin oxidase modified biocathode (BOD@CFs), the resulting hybrid glucose biofuel cell showed open‐circuit voltage and power density values of 0.75 V and 65 μW cm−2, respectively, which remained intact after 3 weeks.