In situ spectroelectrochemical investigation of a biophotoelectrode based on photoreaction centers embedded in a redox hydrogel

The field of biophotoelectrochemistry and its application in biophotovoltaics and biosensors has gained more and more attention in recent years. Knowledge of the redox potentials of the catalytically active protein cofactors in biophotovoltaic devices is crucial for accurate modelling and in discerning the mechanisms of their operation. Here, for the first time, we used spectroelectrochemical methods to investigate thermodynamic parameters of a biophotoelectrode in situ. We determined redox potentials of two elements of the system: the primary electron donor in photosynthetic reaction centers (RCs) of the bacterium Rhodobacter sphaeroides and osmium-complex based redox mediators that are bound to a hydrogel matrix. We observe that the midpoint potential of the primary donor is shifted towards more positive potentials in comparison to literature data for RCs solubilized in buffered water solution, likely due to interaction with the polymer matrix. We also demonstrate that the osmium-complex modified redox polymer efficiently wires the RCs to the electrode, maintaining a high Internal Quantum Efficiency with approximately one electron per two photons generated (IQE=50±12%). Overall, this biophotoelectrode may be attractive for controlling the redox state of the protein when performing other types of experiments, e.g. time resolved absorption or fluorescence measurements, in order to gain insights into kinetic limitations and thereby help in the rational design of bioelectronic devices.