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The quinohaemoprotein alcohol dehydrogenase from Gluconacetobacter xylinus: molecular and catalytic properties

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Abstract

Gluconacetobacter xylinus possesses a constitutive membrane-bound oxidase system for the use of ethanol. Its alcohol dehydrogenase complex (ADH) was purified to homogeneity and characterized. It is a 119-kDa heterodimer (68 and 41 kDa subunits). The peroxidase reaction confirmed the presence of haem C in both subunits. Four cytochromes c per enzyme were determined by pyridine hemochrome spectroscopy. Redox titrations of the purified ADH revealed the presence of four haem c redox centers, with apparent mid-point potential values (Em7) of −33, +55, +132 and +310 mV, respectively. The ADH complex contains one mol of pyrroloquinoline quinone as determined by HPLC. The enzyme was purified in full reduced state; oxidation was induced by potassium ferricyanide and substrate restores full reduction. Activity responses to pH were sharp, showing two distinct optimal pH values (i.e. pH 5.5 and 6.5) depending on the electron acceptor used. Purified ADH oxidizes primary alcohols (C2–C6) but not methanol. Noteworthy, aliphatic aldehydes (C1–C4) were also good substrates. Myxothiazol and antymicin A were powerful inhibitors of the purified ADH complex, most likely acting at the ubiquinone acceptor site in subunit II.

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Acknowledgments

This work was supported in part by grants from Consejo Nacional de Ciencia y Tecnología 50672 and by Universidad Nacional Autónoma de México (Programa de Apoyo a Proyectos de investigación e Innovación Tecnológica UNAM, IN220508). We are grateful to Juan Manuel Barbosa Castillo and Ivette Rosas Arciniega for assistance in computer techniques and María del Rocío Romualdo Martínez for secretarial assistance.

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Authors and Affiliations

  1. Departamento de Bioquímica, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Apdo. Postal 70-242, 04510, México, México

    J. L. Chávez-Pacheco, M. Contreras-Zentella, S. Gómez-Manzo & J. E. Escamilla

  2. Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Apdo. Postal 70-242, 04510, México, México

    G. Mendoza-Hernández

  3. Avenida Copílco 162, Coyoacán, 04314, México, México

    J. Membrillo-Hernández

  4. Departamento de Bioquímica, Instituto de Química, Universidad Nacional Autónoma de México, Apdo. Postal 70-242, 04510, México, México

    R. Arreguín-Espinoza

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  1. J. L. Chávez-Pacheco
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  2. M. Contreras-Zentella
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  5. G. Mendoza-Hernández
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  6. S. Gómez-Manzo
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  7. J. E. Escamilla
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Correspondence to J. E. Escamilla.

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Communicated by Timothy Donohue.

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Chávez-Pacheco, J.L., Contreras-Zentella, M., Membrillo-Hernández, J. et al. The quinohaemoprotein alcohol dehydrogenase from Gluconacetobacter xylinus: molecular and catalytic properties. Arch Microbiol 192, 703–713 (2010). https://doi.org/10.1007/s00203-010-0598-0

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  • DOI: https://doi.org/10.1007/s00203-010-0598-0

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