Biomolecules are inherently less conductive. Therefore, bio-electronic devices that depend on conventional biomolecules to tether enzymes onto electrode supports and to shuttle electrons between the enzyme and the electrode suffer from charge dissipation. This results in bioanondes with decreased current-voltage responses as a result of ohmic losses. Thus, lack of an effective molecular wiring system that can allow unimpeded charge transport is a significant problem that hinders our ability to utilize the full potential of enzymatic self-powering bioelectronic devices. Reducing the internal resistance is the simplest way of increasing the current-voltage response associated with bioanodes and has yet been an unmet challenge. In living cells, iron-sulfur complexes ([Fe-S]) are known to aid in circumventing these issues in the mitochondrial electron transport chains. We describe a technique to attach iron-sulfur moieties to gold surface in non-aqueous media initially and then continue coenzyme and apoenzyme attachment in aqueous media while keeping the efficacy of FeS and the enzyme system intact. As a working model, a glycerol-sensitive gold bioanode is described based on direct attachment of the glycerol-dehydrogenase (GlDH)-NAD⁺ apoenzyme-coenzyme complex onto the supporting gold surface using iron (II) sulfide (FeS) mediation. A conventional pyrroloquinoline quinone (PQQ)-based electrode was used as the control. The performances of the two electrode systems were compared using amperometric and potentiometric studies. Successful tethering of the molecular wiring schemes was verified using cyclic voltammetry and spectroscopy. Amperometric and potentiometric analyses with glycerol dehydrogenase-based model electrodes confirmed the ability of this single-molecule to remarkably amplify, about ten-fold increase in current and up to 24% increase in voltage outputs, as compared to electrodes fabricated with the conventional PQQ-based composite molecular wiring system. FeS achieves the dual purpose of anchoring the enzyme to the gold electrode while also mediating electron shuttling between coenzyme and the electrode surface. This dual functionality allows usage of a single-molecular wire to foster electrical communication between the enzyme and the electrode instead of the conventional multi-molecular wiring system and in turn reducing the internal resistance of the electrode. The resulting increase in current/voltage response opens up a wide range of possibilities for developing efficient bio-electrodes for bioelectronics applications. The importance of this work is the ability to reduce the internal resistance (and thus the overpotential) of circuits that use redox enzymes - allowing complete utilization of the “available power” and “signal capacity” in enzyme-based bioelectronic systems.