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Template‐Synthesized Protein Macroporous Biofilms: Conformational Related Direct Electron Transfer

We report on the synthesis of three‐dimensionally ordered structure (3D) of macroporous cytochrome c (cyt‐c) biofilms by using the inverted colloidal polystyrene crystal template technique and Triton X‐100 as entrapping matrix. Electrochemical impedance spectroscopic (EIS) measurements reveal that the immobilized cyt‐c molecules exhibit fast interfacial electron‐communication rate. We found that the orientation of the cyt‐c molecules entrapped in the 3D macroporous structure was determined by the interfacial electric field. Higher interfacial electric field limits the reorientational flexibility of the entrapped cyt‐c molecules, resulting in lower intermolecular electron‐communication rate constant (k°). Therefore, the highest intermolecular electron‐communication rate constant can only be obtained at potentials approaching to the potential of zero charge of the gold electrode, since k° is mainly determined by the molecular orientation in the biofilm. In addition, the prepared biofilms with enhanced functional density could find potential application in the bioelectronic sensing system.