Bacterial Peroxidase on Electrochemically Reduced Graphene Oxide for Highly Sensitive H₂ O₂ Detection

Peroxidase enzymes enable the construction of electrochemical sensors for highly sensitive and selective quantitative detection of various molecules, pathogens and diseases. Herein, we describe the immobilization of a peroxidase from Bacillus s. (BsDyP) on electrochemically reduced graphene oxide (ERGO) deposited on indium tin oxide (ITO) and polyethylene terephthalate (PET) layers. XRD, SEM, AFM, FT-IR and Raman characterization of the sensor confirmed its structural integrity and a higher enzyme surface occupancy. The BsDyP-ERGO/ITO/PET electrode performed better than other horseradish peroxidase-based electrodes, as evinced by an improved electrochemical response in the nanomolar range (linearity 0.05-280 μM of H₂ O₂ , LOD 32 nM). The bioelectrode was mechanically robust, active in the 3.5-6 pH range and exhibited no loss of activity upon storage for 8 weeks at 4 °C.