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    Immobilization of Alcohol Dehydrogenase in Phospholipid Langmuir−Blodgett Films To Detect Ethanol
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    Departamento de Ciências Exatas e da Terra, Universidade Federal de São Paulo, Diadema, and Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, São Paulo, Brazil, and KSV Instruments Ltd., Helsinki, Finland
    * To whom correspondence should be addressed. E-mail: [email protected]. Phone: (55-11) 4049-3300. Fax: (55-11) 4043-6428.
    †Universidade Federal de São Paulo.
    ‡Universidade de São Paulo.
    §KSV Instruments Ltd.
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    Langmuir

    Cite this: Langmuir 2009, 25, 5, 3057–3061
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    https://doi.org/10.1021/la8037445
    Published February 3, 2009
    Copyright © 2009 American Chemical Society
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    Abstract

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    Enzyme immobilization in nanostructured films may be useful for a number of biomimetic systems, particularly if suitable matrixes are identified. Here we show that alcohol dehydrogenase (ADH) has high affinity toward a negatively charged phospholipid, dimyristoylphosphatidic acid (DMPA), which forms a Langmuir monolayer at an air−water interface. Incorporation of ADH into the DMPA monolayer was monitored with surface pressure measurements and polarization-modulation infrared reflection absorption spectroscopy, with the α-helices from ADH being mainly oriented parallel to the water surface. ADH remained at the interface even at high surface pressures, thus allowing deposition of Langmuir−Blodgett (LB) films from the DMPA−ADH film. Indeed, interaction with DMPA enhances the transfer of ADH, where the mass transferred onto a solid support increased from 134 ng for ADH on a Gibbs monolayer to 178 ng for an LB film with DMPA. With fluorescence spectroscopy it was possible to confirm that the ADH structure was preserved even after one month of the LB deposition. ADH-containing films deposited onto gold-interdigitated electrodes were employed in a sensor array capable of detecting ethanol at concentrations down to 10 ppb (in volume), using impedance spectroscopy as the method of detection.

    Copyright © 2009 American Chemical Society

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    Figure showing the adsorption kinetics for ADH at a DMPA monolayer and tables giving the capacitance data used in PCA calculus and eigenvalue data. This information is available free of charge via the Internet at http://pubs.acs.org.

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    Langmuir

    Cite this: Langmuir 2009, 25, 5, 3057–3061
    Click to copy citationCitation copied!
    https://doi.org/10.1021/la8037445
    Published February 3, 2009
    Copyright © 2009 American Chemical Society
    Request reuse permissions

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