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Purification and Characterisation of a 31-kDa Chitinase from the Myzus Persicae Aphid: A Target for Hemiptera Biocontrol

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Abstract

Hydrolytic enzymes involved in chitin degradation are important to allow moulting during insect development. Chitinases are interesting targets to disturb growth and develop alternative strategies to control insect pests. In this work, a chitinase from the aphid Myzus persicae was purified with a 36-fold purification rate in a three step procedure by ammonium sulphate fractionation, anion-exchange chromatography on a DEAE column and on an affinity Concanavalin A column. The purified chitinase purity assessed by 1D and 2D SDS–PAGE revealed a single band and three spots at 31 kDa, respectively. Chitinases were found to have high homologies with Concanavalins A and B, two chitinase-related proteins, a fungal endochitinase and an aphid acetylhydrolase by peptide identification by Maldi-Tof-Tof. The efficiency of two potent chitinase inhibitors, namely allosamidin and psammaplin A, was tested and showed significant rate of enzymatic inhibition.

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Acknowledgments

We are grateful to Dr. S. Sakuda (Department of Applied Biological Chemistry, The University of Tokyo, Japan) and Dr. J.N. Tabudravu (Marine Natural Products Laboratory, Department of Chemistry, University of Aberdeen, Scotland, UK) for providing respectively allosamidin and psammaplin A. We thank the ‘Service Régional de la Protection des Végétaux du Nord Pas-de-Calais’ for providing aphids and the ‘Comité Nord Plants de Pommes de Terre’ for providing potato tubers. This work was supported by the Ministère Français de la Recherche, the Conseil Régional de Picardie, the Fonds Social Européen and also the Fond National pour la Recherche Scientifique from Belgium (FNRS) for its funding (FRFC project number 2.4561.06).

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Authors and Affiliations

  1. Entomologie Fonctionnelle et Evolutive, Gembloux Agro-Bio Tech, Université de Liège, Passage des Déportés 2, Gembloux, Liège, Belgium

    Frédéric Francis, Sophie Vandermoten & Eric Haubruge

  2. UPRES EA 3900, Biologie des Plantes et Contrôle des Insectes Ravageurs, Université de Picardie Jules Verne, 33 Rue Saint Leu, 80039, Amiens Cedex, France

    Julien Saguez, Anas Cherqui & Philippe Giordanengo

  3. Centre de Recherche et de Développement en Horticulture, Agriculture et Agro-alimentaire Canada, 430, Boulevard Gouin, Saint-Jean-sur-Richelieu, Québec, J3B 3E6, Canada

    Julien Saguez & Charles Vincent

  4. Biologie Moléculaire et Biotechnologie Végétales, Université de Liège, Boulevard du Rectorat 27, 4000, Liège, Belgium

    Marie-France Versali & Jacques Dommès

  5. Laboratoire de Spectroscopie de Masse, Université de Liège, Allée de la Chimie 3, 4000, Liège, Belgium

    Edwin De Pauw

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  1. Frédéric Francis
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  2. Julien Saguez
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Correspondence to Frédéric Francis.

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Francis, F., Saguez, J., Cherqui, A. et al. Purification and Characterisation of a 31-kDa Chitinase from the Myzus Persicae Aphid: A Target for Hemiptera Biocontrol. Appl Biochem Biotechnol 166, 1291–1300 (2012). https://doi.org/10.1007/s12010-011-9517-3

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  • DOI: https://doi.org/10.1007/s12010-011-9517-3

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