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Spectroscopic and Electrochemical Characterization of Cytochrome P450st‐DDAB Films on a Plastic‐Formed Carbon Electrode
Authors:Supranee Wiwatchaiwong  Hirotoshi Matsumura  Nobuhumi Nakamura  Masafumi Yohda  Hiroyuki Ohno
Institution:Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184‐8588, Japan
Abstract:We have studied the characterization of thermophilic cytochrome P450 (P450st)‐didodecyldimethylammonium bromide (DDAB) films by using UV‐vis absorption, resonance Raman spectroscopy, and electrochemical methods. The observed Raman spectrum indicated near‐native conformation of the heme iron in DDAB film on the surface of a glass slide, while on the surface of a plastic‐formed carbon (PFC) electrode, the conformation of P450st‐DDAB was very similar to that of heme‐DDAB film, suggesting the release of heme from P450st in DDAB films on PFC electrodes. When NaBr was added as salt to the casting solution, the result of Raman spectrum indicated near‐native conformation of P450st in DDAB film even on the PFC electrode, but no redox potential of P450st which has near native structure was observed. This study suggests the essential experimental conditions when working with heme protein‐DDAB films as, in some cases, heme iron from proteins is released on the surface of the electrode.
Keywords:Surfactant films  Cytochrome P450  Protein film voltammetry  Resonance Raman spectroscopy  Plastic‐formed carbon electrode
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