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Proteomic identification of CBM37-containing cellulases produced by the rumen cellulolytic bacterium Ruminococcus albus 20 and their putative involvement in bacterial adhesion to cellulose

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Abstract

The objective of this study was to identify and characterize other proteins than fimbrial proteins potentially involved in R. albus 20 adhesion to cellulose using an adhesion-related antiserum preparation (i.e. anti-Adh serum). From protein fractions of R. albus 20 grown on cellulose, the serum recognized at least 10 cellulose-binding proteins (CBPs), among which homologs of glycoside hydrolases (family 5, 9 and 48) of R. albus 8 (i.e. Cel5G, Cel9B and Cel48A) were identified by a proteomic approach. In strain 20, Cel9B and Cel48A were identified as two major CBPs and as bacterial cell-associated proteins. The anti-Adh serum was also shown to target the C-terminal family 37 carbohydrate-binding module (CBM37) of Cel9B and Cel48A, indicating that this module, unique to R. albus, may play a significant role in bacterial adhesion to cellulose as suggested previously for R. albus 8. Overall, our results support the hypothesis of an adhesion mechanism involving the CBM37 of Cel9B and Cel48A. This adhesion mechanism may not be restricted to these two enzymes but may also involve other CBM37-containing proteins such as Cel5G and the other uncharacterised proteins recognized by the anti-Adh serum.

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Abbreviations

2-DE:

Two-dimensional gel electrophoresis

IEF:

Isoelectric focusing

SDS-PAGE:

Sodium dodecyl sulfate poly-acrylamide gel electrophoresis

CBP:

Cellulose-binding protein

CBM37:

Family 37 carbohydrate-binding module

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Acknowledgments

We are very grateful to Y. Ribot and G. Andant for technical assistance in DNA sequencing and 2-DE protein analysis, respectively.

Author information

Author notes

  1. Harivony Rakotoarivonina

    Present address: INRA, Université de Reims, FARE UMR614, 51688, Reims, France

Authors and Affiliations

  1. INRA, UR454 Unité de Microbiologie, 63122, Saint-Genès-Champanelle, France

    Harivony Rakotoarivonina, Cécile Terrie, Evelyne Forano & Pascale Mosoni

  2. INRA Plate Forme de Protéomique, 63122, Saint-Genès-Champanelle, France

    Christophe Chambon

Authors
  1. Harivony Rakotoarivonina
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  2. Cécile Terrie
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  3. Christophe Chambon
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  4. Evelyne Forano
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  5. Pascale Mosoni
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Corresponding author

Correspondence to Pascale Mosoni.

Additional information

Communicated by Wolfgang Buckel.

The EMBL accession numbers for the sequences reported in this paper are FM872295 for Cel9B and FM872296 for Cel48A.

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Rakotoarivonina, H., Terrie, C., Chambon, C. et al. Proteomic identification of CBM37-containing cellulases produced by the rumen cellulolytic bacterium Ruminococcus albus 20 and their putative involvement in bacterial adhesion to cellulose. Arch Microbiol 191, 379–388 (2009). https://doi.org/10.1007/s00203-009-0463-1

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  • DOI: https://doi.org/10.1007/s00203-009-0463-1

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