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The carbohydrate-binding module of Fragaria × ananassa expansin 2 (CBM-FaExp2) binds to cell wall polysaccharides and decreases cell wall enzyme activities “in vitro”

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Abstract

A putative carbohydrate binding module (CBM) from strawberry (Fragaria × ananassa Duch.) expansin 2 (CBM-FaExp2) was cloned and the encoding protein was over-expressed in Escherichia coli and purified in order to evaluate its capacity to bind different cell wall polysaccharides “in vitro”. The protein CBM-FaExp2 bound to microcrystalline cellulose, xylan and pectin with different affinities (Kad = 33.6 ± 0.44 mL g−1, Kad = 11.37 ± 0.87 mL g−1, Kad = 10.4 ± 0.19 mL g−1, respectively). According to “in vitro” enzyme assays, this CBM is able to decrease the activity of cell wall degrading enzymes such as polygalacturonase, endo-glucanase, pectinase and xylanase, probably because the binding of CBM-FaExp2 to the different substrates interferes with enzyme activity. The results suggest that expansins would bind not only cellulose but also a wide range of cell wall polymers.

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Acknowledgments

This work has been supported by grants from CONICET (PIP 0062), ANPCYT (PICT 2006-01140) and UNSAM (Biotechnology Program 2007). Authors thank José Luis Burgos (Comisión de Investigaciones Científicas, CIC) for valuable technical assistance.

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Authors and Affiliations

  1. IIB-INTECH (CONICET-UNSAM), Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús, Camino de Circunvalación Laguna, km 6, (B7130IWA) Chascomús, Pcia, Buenos Aires, Argentina

    Cristina Nardi, Cristian Escudero, Natalia Villarreal & Gustavo Martínez

  2. Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP), 47 y 115, 1900, La Plata, Argentina

    Gustavo Martínez & Pedro Marcos Civello

  3. INFIVE (CONICET-UNLP), Instituto de Fisiología Vegetal, Diag, 113 y Calle 61, n° 495-C.c 327, 1900, La Plata, Argentina

    Pedro Marcos Civello

Authors
  1. Cristina Nardi
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  2. Cristian Escudero
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  3. Natalia Villarreal
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  4. Gustavo Martínez
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  5. Pedro Marcos Civello
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Correspondence to Pedro Marcos Civello.

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C. Nardi and C. Escudero contributed equally to this work.

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Nardi, C., Escudero, C., Villarreal, N. et al. The carbohydrate-binding module of Fragaria × ananassa expansin 2 (CBM-FaExp2) binds to cell wall polysaccharides and decreases cell wall enzyme activities “in vitro”. J Plant Res 126, 151–159 (2013). https://doi.org/10.1007/s10265-012-0504-8

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