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Journal of Bacteriology, July 2007, p. 4774-4783, Vol. 189, No. 13
0021-9193/07/$08.00+0     doi:10.1128/JB.00143-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

A Novel Cell Surface-Anchored Cellulose-Binding Protein Encoded by the sca Gene Cluster of Ruminococcus flavefaciens{triangledown}

Marco T. Rincon,1 Tadej Cepeljnik,2 Jennifer C. Martin,1 Yoav Barak,4 Raphael Lamed,3 Edward A. Bayer,4 and Harry J. Flint1*

Microbial Ecology Group, Rowett Research Institute, Aberdeen, United Kingdom,1 Zootechnical Department, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia,2 Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Ramat Aviv, Israel,3 Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel4

Received 29 January 2007/ Accepted 19 April 2007

Ruminococcus flavefaciens produces a cellulosomal enzyme complex, based on the structural proteins ScaA, -B, and -C, that was recently shown to attach to the bacterial cell surface via the wall-anchored protein ScaE. ScaA, -B, -C, and -E are all cohesin-bearing proteins encoded by linked genes in the sca cluster. The product of an unknown open reading frame within the sca cluster, herein designated CttA, is similar in sequence at its C terminus to the corresponding region of ScaB, which contains an X module together with a dockerin sequence. The ScaB-XDoc dyad was shown previously to interact tenaciously with the cohesin of ScaE. Likewise, avid binding was confirmed between purified recombinant fragments of the CttA-XDoc dyad and the ScaE cohesin. In addition, the N-terminal regions of CttA were shown to bind to cellulose, thus suggesting that CttA is a cell wall-anchored, cellulose-binding protein. Proteomic analysis showed that the native CttA protein (~130 kDa) corresponds to one of the three most abundant polypeptides binding tightly to insoluble cellulose in cellulose-grown R. flavefaciens 17 cultures. Interestingly, this protein was also detected among cellulose-bound proteins in the related strain R. flavefaciens 007C but not in a mutant derivative, 007S, that was previously shown to have lost the ability to grow on dewaxed cotton fibers. In R. flavefaciens, the presence of CttA on the cell surface is likely to provide an important mechanism for substrate binding, perhaps compensating for the absence of an identified cellulose-binding module in the major cellulosomal scaffolding proteins of this species.


* Corresponding author. Mailing address: The Rowett Research Institute, Microbial Ecology Group, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, United Kingdom. Phone: 44-(0)1224-712751. Fax: 44-(0)1224-716687. E-mail: H.Flint{at}rowett.ac.uk

{triangledown} Published ahead of print on 27 April 2007.


Journal of Bacteriology, July 2007, p. 4774-4783, Vol. 189, No. 13
0021-9193/07/$08.00+0     doi:10.1128/JB.00143-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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