Renewable dehydrogenase-based interfaces for bioelectronic applications

Brian L. Hassler, Neeraj Kohli, J. Gregory Zeikus, Ilsoon Lee, Robert M. Worden

    Research output: Contribution to journalArticle

    • 31 Citations

    Abstract

    Bioelectronic interfaces that establish electrical communication between redox enzymes and electrodes have potential applications as biosensors, biocatalytic reactors, and biological fuel cells. However, these interfaces contain labile components, including enzymes and cofactors, which have limited lifetimes and must be replaced periodically to allow long-term operation. Current methods to fabricate bioelectronic interfaces do not allow facile replacement of these components, thus limiting the useful lifetime of the interfaces. In this paper we describe a versatile new fabrication approach that binds the enzymes and cofactors using reversible ionic interactions. This approach allows the interface to be removed via a simple pH change and then replaced to fully regenerate the biocatalytic activity. The positively charged polyelectrolyte poly(ethylenimine) was used to ionically bond a dehydrogenase enzyme and its cofactor to a gold electrode that was functionalized with 3-mercaptopropionic acid and the electron mediator toluidine blue O. By reducing the pH, the surface-bound 3-mercaptopropionic acid was protonated, disrupting the ionic bonds and releasing the enzyme-modified polyelectrolyte. After neutralization, fresh enzyme and cofactor were bound, regenerating the bioelectronic interface. Cyclic voltammetry, chronoamperometry, constant potential amperometry, electrochemical impedance spectroscopy, and Fourier transform infrared spectroscopy analyses were used to characterize the bioelectronic interfaces. For the two enzymes tested (secondary alcohol dehydrogenase and sorbitol dehydrogenase) and their respective cofactors (β-nicotinamide adenine dinucleotide phosphate and β-nicotinamide adenine dinucleotide), the reconstituted interface exhibited a surface coverage, an electron-transfer coefficient, and a turnover rate similar to those of the original interface.

    Original languageEnglish (US)
    Pages (from-to)7127-7133
    Number of pages7
    JournalLangmuir
    Volume23
    Issue number13
    DOIs
    StatePublished - Jun 19 2007

    Profile

    Enzymes
    enzymes
    Edema Disease of Swine
    Magnesium Deficiency
    Autoradiography
    Electrons
    dehydrogenases
    Polyelectrolytes
    Electrodes
    Acids
    Carubicin
    beta-Amylase
    Delivery Rooms
    Ethane
    Electrochemical impedance spectroscopy
    Biosensors
    Oxidation-Reduction
    Communication
    nicotinamide
    adenines

    ASJC Scopus subject areas

    • Physical and Theoretical Chemistry
    • Colloid and Surface Chemistry

    Cite this

    Renewable dehydrogenase-based interfaces for bioelectronic applications. / Hassler, Brian L.; Kohli, Neeraj; Zeikus, J. Gregory; Lee, Ilsoon; Worden, Robert M.

    In: Langmuir, Vol. 23, No. 13, 19.06.2007, p. 7127-7133.

    Research output: Contribution to journalArticle

    Hassler, BL, Kohli, N, Zeikus, JG, Lee, I & Worden, RM 2007, 'Renewable dehydrogenase-based interfaces for bioelectronic applications' Langmuir, vol 23, no. 13, pp. 7127-7133. DOI: 10.1021/la7004437
    Hassler BL, Kohli N, Zeikus JG, Lee I, Worden RM. Renewable dehydrogenase-based interfaces for bioelectronic applications. Langmuir. 2007 Jun 19;23(13):7127-7133. Available from, DOI: 10.1021/la7004437

    Hassler, Brian L.; Kohli, Neeraj; Zeikus, J. Gregory; Lee, Ilsoon; Worden, Robert M. / Renewable dehydrogenase-based interfaces for bioelectronic applications.

    In: Langmuir, Vol. 23, No. 13, 19.06.2007, p. 7127-7133.

    Research output: Contribution to journalArticle

    @article{44d0b8a85423471d8a1ea0c60470fc45,
    title = "Renewable dehydrogenase-based interfaces for bioelectronic applications",
    abstract = "Bioelectronic interfaces that establish electrical communication between redox enzymes and electrodes have potential applications as biosensors, biocatalytic reactors, and biological fuel cells. However, these interfaces contain labile components, including enzymes and cofactors, which have limited lifetimes and must be replaced periodically to allow long-term operation. Current methods to fabricate bioelectronic interfaces do not allow facile replacement of these components, thus limiting the useful lifetime of the interfaces. In this paper we describe a versatile new fabrication approach that binds the enzymes and cofactors using reversible ionic interactions. This approach allows the interface to be removed via a simple pH change and then replaced to fully regenerate the biocatalytic activity. The positively charged polyelectrolyte poly(ethylenimine) was used to ionically bond a dehydrogenase enzyme and its cofactor to a gold electrode that was functionalized with 3-mercaptopropionic acid and the electron mediator toluidine blue O. By reducing the pH, the surface-bound 3-mercaptopropionic acid was protonated, disrupting the ionic bonds and releasing the enzyme-modified polyelectrolyte. After neutralization, fresh enzyme and cofactor were bound, regenerating the bioelectronic interface. Cyclic voltammetry, chronoamperometry, constant potential amperometry, electrochemical impedance spectroscopy, and Fourier transform infrared spectroscopy analyses were used to characterize the bioelectronic interfaces. For the two enzymes tested (secondary alcohol dehydrogenase and sorbitol dehydrogenase) and their respective cofactors (β-nicotinamide adenine dinucleotide phosphate and β-nicotinamide adenine dinucleotide), the reconstituted interface exhibited a surface coverage, an electron-transfer coefficient, and a turnover rate similar to those of the original interface.",
    author = "Hassler, {Brian L.} and Neeraj Kohli and Zeikus, {J. Gregory} and Ilsoon Lee and Worden, {Robert M.}",
    year = "2007",
    month = "6",
    doi = "10.1021/la7004437",
    volume = "23",
    pages = "7127--7133",
    journal = "Langmuir",
    issn = "0743-7463",
    publisher = "American Chemical Society",
    number = "13",

    }

    TY - JOUR

    T1 - Renewable dehydrogenase-based interfaces for bioelectronic applications

    AU - Hassler,Brian L.

    AU - Kohli,Neeraj

    AU - Zeikus,J. Gregory

    AU - Lee,Ilsoon

    AU - Worden,Robert M.

    PY - 2007/6/19

    Y1 - 2007/6/19

    N2 - Bioelectronic interfaces that establish electrical communication between redox enzymes and electrodes have potential applications as biosensors, biocatalytic reactors, and biological fuel cells. However, these interfaces contain labile components, including enzymes and cofactors, which have limited lifetimes and must be replaced periodically to allow long-term operation. Current methods to fabricate bioelectronic interfaces do not allow facile replacement of these components, thus limiting the useful lifetime of the interfaces. In this paper we describe a versatile new fabrication approach that binds the enzymes and cofactors using reversible ionic interactions. This approach allows the interface to be removed via a simple pH change and then replaced to fully regenerate the biocatalytic activity. The positively charged polyelectrolyte poly(ethylenimine) was used to ionically bond a dehydrogenase enzyme and its cofactor to a gold electrode that was functionalized with 3-mercaptopropionic acid and the electron mediator toluidine blue O. By reducing the pH, the surface-bound 3-mercaptopropionic acid was protonated, disrupting the ionic bonds and releasing the enzyme-modified polyelectrolyte. After neutralization, fresh enzyme and cofactor were bound, regenerating the bioelectronic interface. Cyclic voltammetry, chronoamperometry, constant potential amperometry, electrochemical impedance spectroscopy, and Fourier transform infrared spectroscopy analyses were used to characterize the bioelectronic interfaces. For the two enzymes tested (secondary alcohol dehydrogenase and sorbitol dehydrogenase) and their respective cofactors (β-nicotinamide adenine dinucleotide phosphate and β-nicotinamide adenine dinucleotide), the reconstituted interface exhibited a surface coverage, an electron-transfer coefficient, and a turnover rate similar to those of the original interface.

    AB - Bioelectronic interfaces that establish electrical communication between redox enzymes and electrodes have potential applications as biosensors, biocatalytic reactors, and biological fuel cells. However, these interfaces contain labile components, including enzymes and cofactors, which have limited lifetimes and must be replaced periodically to allow long-term operation. Current methods to fabricate bioelectronic interfaces do not allow facile replacement of these components, thus limiting the useful lifetime of the interfaces. In this paper we describe a versatile new fabrication approach that binds the enzymes and cofactors using reversible ionic interactions. This approach allows the interface to be removed via a simple pH change and then replaced to fully regenerate the biocatalytic activity. The positively charged polyelectrolyte poly(ethylenimine) was used to ionically bond a dehydrogenase enzyme and its cofactor to a gold electrode that was functionalized with 3-mercaptopropionic acid and the electron mediator toluidine blue O. By reducing the pH, the surface-bound 3-mercaptopropionic acid was protonated, disrupting the ionic bonds and releasing the enzyme-modified polyelectrolyte. After neutralization, fresh enzyme and cofactor were bound, regenerating the bioelectronic interface. Cyclic voltammetry, chronoamperometry, constant potential amperometry, electrochemical impedance spectroscopy, and Fourier transform infrared spectroscopy analyses were used to characterize the bioelectronic interfaces. For the two enzymes tested (secondary alcohol dehydrogenase and sorbitol dehydrogenase) and their respective cofactors (β-nicotinamide adenine dinucleotide phosphate and β-nicotinamide adenine dinucleotide), the reconstituted interface exhibited a surface coverage, an electron-transfer coefficient, and a turnover rate similar to those of the original interface.

    UR - http://www.scopus.com/inward/record.url?scp=34547244184&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=34547244184&partnerID=8YFLogxK

    U2 - 10.1021/la7004437

    DO - 10.1021/la7004437

    M3 - Article

    VL - 23

    SP - 7127

    EP - 7133

    JO - Langmuir

    T2 - Langmuir

    JF - Langmuir

    SN - 0743-7463

    IS - 13

    ER -