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Characterization of structure and activity of garlic peroxidase (POX1B)

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Abstract

Structural characterization and study of the activity of new POX1B protein from garlic which has a high peroxidase activity and can be used as a biosensor for the detection of hydrogen peroxide and phenolic compounds were performed and compared with the findings for other heme peroxidases. The structure–function relationship was investigated by analysis of the spectroscopic properties and correlated to the structure determined by a new generation of high-performance hybrid mass spectrometers. The reactivity of the enzyme was analyzed by studies of the redox activity toward various ligands and the reactivity with various substrates. We demonstrated that, in the case of garlic peroxidase, the heme group is pentacoordinated, and has an histidine as a proximal ligand. POX1B exhibited a high affinity for hydrogen peroxide as well as various reducing cosubstrates. In addition, high enzyme specificity was demonstrated. The k cat and K M values were 411 and 400 mM−1 s−1 for 3,3′,5,5′-tetramethylbenzidine and 2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), respectively. Furthermore, the reduction of nitro compounds in the presence of POX1B was demonstrated by iron(II) nitrosoalkane complex assay. In addition, POX1B showed a great potential for application for drug metabolism since its ability to react with 1-nitrohexane in the presence of sodium dithionite was demonstrated by the appearance of a characteristic Soret band at 411 nm. The high catalytic efficiency obtained in the case of the new garlic peroxidase (POX1B) is suitable for the monitoring of different analytes and biocatalysis.

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Acknowledgments

This work was supported by a cooperation project between the CNRS (France) and DGRSRT (Tunisia). Financial support from the Tunisian Ministry of High Education, Scientific Research and Technology and Paris-Sud 11 University is gratefully acknowledged.

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Authors and Affiliations

  1. Equipe de Chimie Bioorganique et Bioinorganique, Institut de Chimie Moléculaire et Matériaux d’Orsay, UMR8182, CNRS, Université Paris-Sud, 91405, Orsay, France

    Sarra El Ichi, Anna Miodek, Hélène Sauriat-Dorizon, Jean-Pierre Mahy & Hafsa Korri-Youssoufi

  2. Bioengineering Unit 99 UR 09-26, National Institute of Applied Sciences and Technology, 7th November University at Carthage, Centre Urbain Nord, BP 676-1080, Tunis Cedex, Tunisia

    Sarra El Ichi & Mohamed Nejib Marzouki

  3. Unité BioBac, INRA UR477, Jouy-en-Josas, France

    Céline Henry

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  1. Sarra El Ichi
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  2. Anna Miodek
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  3. Hélène Sauriat-Dorizon
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  4. Jean-Pierre Mahy
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Correspondence to Hafsa Korri-Youssoufi.

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El Ichi, S., Miodek, A., Sauriat-Dorizon, H. et al. Characterization of structure and activity of garlic peroxidase (POX1B). J Biol Inorg Chem 16, 157–172 (2011). https://doi.org/10.1007/s00775-010-0714-2

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