Amperometric hydrogen peroxide biosensor based on the immobilization of heme proteins on gold nanoparticles–bacteria cellulose nanofibers nanocomposite

A novel matrix, gold nanoparticles–bacterial cellulose nanofibers (Au–BC) nanocomposite was developed for enzyme immobilization and biosensor fabrication due to its unique properties such as satisfying biocompatibility, good conductivity and extensive surface area, which were inherited from both gold nanoparticles (AuNPs) and bacterial cellulose nanofibers (BC). Heme proteins such as horseradish peroxidase (HRP), hemoglobin (Hb) and myoglobin (Mb) were successfully immobilized on the surface of Au–BC nanocomposite modified glassy carbon electrode (GCE). The immobilized heme proteins showed electrocatalytic activities to the reduction of H2O2 in the presence of the mediator hydroquinone (HQ), which might be due to the fact that heme proteins retained the near-native secondary structures in the Au–BC nanocomposite which was proved by UV–vis and IR spectra. The response of the developed biosensor to H2O2 was related to the amount of AuNPs in Au–BC nanocomposite, indicating that the AuNPs in BC network played an important role in the biosensor performance. Under the optimum conditions, the biosensor based on HRP exhibited a fast amperometric response (within 1 s) to H2O2, a good linear response over a wide range of concentration from 0.3 μM to 1.00 mM, and a low detection limit of 0.1 μM based on S/N = 3. The high performance of the biosensor made Au–BC nanocomposite superior to other materials as immobilization matrix.