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Electroactive cytochromeP450BM3 cast polyion films on graphite electrodes

Abstract : Films of electrochemically active cytochrome P450BM3 were constructed on graphite electrodes using alternate assembly with polyethyleneimin(PEI). The original layer-by-layer adsorption method was slightly modified here to form so-called “cast polyion” films. The cast polyion films were elaborated by immobilizing two successive layers of PEI and protein in very large excess with respect to a monolayer, without any intermediate washing step. Following the immobilization steps by SEM showed that uniform films of a few micrometers were deposited on the graphite surface. The electrochemically activity of the immobilized cytP450 was tested with regard to the reduction of oxygen and the one-electron reduction of the heme. Cyclic voltammetry indicated surface concentration of electrochemically active cytP450 around 0.6 nmol/cm2, which corresponded to 5% of the total amount of protein that was consumed by the immobilisation process. Adapting the procedure to a graphite felt electrode with the view of scaling up porous electrodes for large scale synthesis increased the concentration to 0.9 nmol/cm2. Cast polyion films may represent a simple technique to immobilize high amount of electrochemically active protein, keeping the advantage of the electrostatic interactions of the regular layer-by-layer method.
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Submitted on : Friday, April 23, 2010 - 1:59:59 PM
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  • HAL Id : hal-00475995, version 1



Aurélie Pardo-Jacques, Régine Basséguy, Alain Bergel. Electroactive cytochromeP450BM3 cast polyion films on graphite electrodes. Electrochimica Acta, Elsevier, 2006, vol. 52 n° 3., 979-987 available on : ⟨hal-00475995⟩



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