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Stationary photocurrent generation from bacteriorhodopsin-loaded lipo-polymersomes in polyelectrolyte multilayer assembly on polyethersulfone membrane

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Abstract

Vesicles constructed of either synthetic polymers alone (polymersomes) or a combination of polymers and lipids (lipo-polymersomes) demonstrate excellent long-term stability and ability to integrate membrane proteins. Applications using lipo-polymersomes with integrated membrane proteins require suitable supports to maintain protein functionality. Using lipo-polymersomes loaded with the light-driven proton pump bacteriorhodopsin (BR), we demonstrate here how the photocurrent is influenced by a chosen support. In our study, we deposited BR-loaded lipo-polymersomes in a cross-linked polyelectrolyte multilayer assembly either directly physisorbed on gold electrode microchips or cross-linked on an intermediary polyethersulfone (PES) membrane covalently grafted using a hydrogel cushion. In both cases, electrochemical impedance spectroscopic characterization demonstrated successful polyelectrolyte assembly with BR-loaded lipo-polymersomes. Light-induced proton pumping by BR-loaded lipo-polymersomes in the different support constructs was characterized by amperometric recording of the generated photocurrent. Application of the hydrogel/PES membrane support together with the polyelectrolyte assembly decreased the transient current response upon light activation of BR, while enhancing the generated stationary current to over 700 nA/cm2. On the other hand, the current response from BR-loaded lipo-polymersomes in a polyelectrolyte assembly without the hydrogel/PES membrane support was primarily a transient peak combined with a low-nanoampere-level stationary photocurrent. Hence, the obtained results demonstrated that by using a hydrogel/PES support it was feasible to monitor continuously light-induced proton flux in biomimetic applications of lipo-polymersomes.

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Acknowledgments

This work was financially supported by the Copenhagen Research School of Nanotechnology (CONT) in Denmark, and the cleantech company Aquaporin A/S in Copenhagen, Denmark. Nanna Bild from DTU Healthtech is acknowledged for the artwork in Fig. 1.

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  1. Agnieszka Mech-Dorosz

    Present address: Novo Nordisk A/S, Brennum Park 24 K, 3400, Hillerød, Denmark

Authors and Affiliations

  1. Department of Biotechnology and Biomedicine, Technical University of Denmark, Produktionstorvet, Building 423, 2800, Kgs. Lyngby, Denmark

    Agnieszka Mech-Dorosz, Jenny Emnéus & Arto Heiskanen

  2. Department of Environmental Engineering, Technical University of Denmark, Bygningstorvet, Building 115, 2800, Kgs. Lyngby, Denmark

    Niada Bajraktari & Claus Hélix-Nielsen

  3. Aquaporin A/S, Nymøllevej 78, 2800, Kgs. Lyngby, Denmark

    Niada Bajraktari & Claus Hélix-Nielsen

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  1. Agnieszka Mech-Dorosz
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Mech-Dorosz, A., Bajraktari, N., Hélix-Nielsen, C. et al. Stationary photocurrent generation from bacteriorhodopsin-loaded lipo-polymersomes in polyelectrolyte multilayer assembly on polyethersulfone membrane. Anal Bioanal Chem 412, 6307–6318 (2020). https://doi.org/10.1007/s00216-020-02533-8

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