Potentiation of the synergistic activities of chitinases ChiA, ChiB and ChiC from Serratia marcescens CFFSUR-B2 by chitobiase (Chb) and chitin binding protein (CBP)
Por:
Gutiérrez-Román M.I., Dunn M.F., Tinoco-Valencia R., Holguín-Meléndez F., Huerta-Palacios G., Guillén-Navarro K.
Publicada:
1 ene 2014
Resumen:
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. © 2013 Springer Science+Business Media Dordrecht.
Filiaciones:
Gutiérrez-Román M.I.:
Departamento de Agricultura, Sociedad y Ambiente, El Colegio de la Frontera Sur, Carretera Antiguo Aeropuerto Km. 2.5, C.P. 30700 Tapachula, Chiapas, Mexico
Dunn M.F.:
UNAM, Programa Genom Func Procariotes, Ctr Ciencias Genom, Cuernavaca 62210, Morelos, Mexico
Tinoco-Valencia R.:
UNAM, Inst Biotecnol, Cuernavaca 62210, Morelos, Mexico
Holguín-Meléndez F.:
Laboratorio de Diagnóstico Fitosanitario, El Colegio de la Frontera Sur, Carretera Antiguo Aeropuerto Km. 2.5, 30700 Tapachula, Chiapas, Mexico
Huerta-Palacios G.:
Departamento de Agricultura, Sociedad y Ambiente, El Colegio de la Frontera Sur, Carretera Antiguo Aeropuerto Km. 2.5, C.P. 30700 Tapachula, Chiapas, Mexico
Guillén-Navarro K.:
Laboratorio de Biotecnología Ambiental y Agroecológica, El Colegio de la Frontera Sur, Carretera Antiguo Aeropuerto Km. 2.5, C.P. 30700 Tapachula, Chiapas, Mexico
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