Potentiation of the synergistic activities of chitinases ChiA, ChiB and ChiC from Serratia marcescens CFFSUR-B2 by chitobiase (Chb) and chitin binding protein (CBP)

  • Martha Ingrid Gutiérrez-Román
  • Michael F. Dunn
  • Raunel Tinoco-Valencia
  • Francisco Holguín-Meléndez
  • Graciela Huerta-Palacios
  • Karina Guillén-Navarro
Original Paper

Abstract

With the goal of understanding the chitinolytic mechanism of the potential biological control strain Serratia marcescens CFFSUR-B2, genes encoding chitinases ChiA, ChiB and ChiC, chitobiase (Chb) and chitin binding protein (CBP) were cloned, the protein products overexpressed in Escherichia coli as 6His-Sumo fusion proteins and purified by affinity chromatography. Following affinity tag removal, the chitinolytic activity of the recombinant proteins was evaluated individually and in combination using colloidal chitin as substrate. ChiB and ChiC were highly active while ChiA was inactive. Reactions containing both ChiB and ChiC showed significantly increased N-acetylglucosamine trimer and dimer formation, but decreased monomer formation, compared to reactions with either enzyme alone. This suggests that while both ChiB and ChiC have a general affinity for the same substrate, they attack different sites and together degrade chitin more efficiently than either enzyme separately. Chb and CBP in combination with ChiB and ChiC (individually or together) increased their chitinase activity. We report for the first time the potentiating effect of Chb on the activity of the chitinases and the synergistic activity of a mixture of all five proteins (the three chitinases, Chb and CBP). These results contribute to our understanding of the mechanism of action of the chitinases produced by strain CFFSUR-B2 and provide a molecular basis for its high potential as a biocontrol agent against fungal pathogens.

Keywords

Chitinases Chitin hydrolysis Serratia marcescens N-acetylglucosamine Chitinolytic activity Synergism 

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Martha Ingrid Gutiérrez-Román
    • 1
  • Michael F. Dunn
    • 2
  • Raunel Tinoco-Valencia
    • 3
  • Francisco Holguín-Meléndez
    • 4
  • Graciela Huerta-Palacios
    • 1
  • Karina Guillén-Navarro
    • 5
  1. 1.Departamento de Agricultura, Sociedad y AmbienteEl Colegio de la Frontera SurTapachulaMexico
  2. 2.Programa de Genómica Funcional de ProcariotesCentro de Ciencias Genómicas UNAMCuernavacaMexico
  3. 3.Instituto de Biotecnología, UNAMCuernavacaMexico
  4. 4.Laboratorio de Diagnóstico FitosanitarioEl Colegio de la Frontera SurTapachulaMexico
  5. 5.Laboratorio de Biotecnología Ambiental y AgroecológicaEl Colegio de la Frontera SurTapachulaMexico

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