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Electroactive cytochromeP450BM3 cast polyion films on graphite electrodes

Pardo-Jacques, Aurélie and Basséguy, Régine and Bergel, Alain Electroactive cytochromeP450BM3 cast polyion films on graphite electrodes. (2006) Electrochimica Acta, vol. 5 (n° 3). pp. 979-987. ISSN 0013-4686

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Official URL: http://dx.doi.org/10.1016/j.electacta.2006.06.035

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.

Item Type:Article
Additional Information:Thanks to Elsevier editor. The definitive version is available at http://www.sciencedirect.com The original PDF of the article can be found at Electrochimica Acta website : http://www.sciencedirect.com/science/journal/00134686
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution: Université de Toulouse > Institut National Polytechnique de Toulouse - INPT
Université de Toulouse > Université Paul Sabatier-Toulouse III - UPS
French research institutions > Centre National de la Recherche Scientifique - CNRS
Laboratory name:
Laboratoire de Génie Chimique - LGC (Toulouse, France) - Bioprocédés et systèmes microbiens (BioSyM)
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Deposited By:Hélène Dubernard

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