A novel composite film based on carbon nanodots (CNDs) and chitosan was readily prepared and used as immobilization matrix to entrap a heme protein, hemoglobin (Hb) for direct electrochemistry and bioelectrocatalysis. A modified electrode was obtained by casting Hb–CNDs–chitosan composites on the glassy carbon (GC) electrode surface. Spectroscopic and electrochemical studies showed that Hb entrapped in the composite film remained in its native structures, and CNDs in the film can greatly facilitate DET between the protein and the GC electrode. The electron-transfer kinetics of Hb in composite film was qualitatively evaluated by using the Marcus theory, and the apparent heterogeneous electron-transfer rate constant (ks) was estimated to be 2.39(±0.03)s−1 with Laviron equations. The modified electrode showed excellent electrocatalytic behavior to the substrate, hydrogen peroxide (H2O2). The linear current response for H2O2 was from 1×10−6 to 1.18×10−4M with a detection limit of 0.27(±0.02)μM at the signal-to-noise ratio of 3, and the apparent Michaelis–Menten constant was 0.067(±0.02)mM. These important features of CNDs–chitosan film have implied to be a promising platform for elaborating bioelectrochemical devices such as biosensors and biofuel cells.
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