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Journal of Bacteriology, November 2005, p. 7569-7578, Vol. 187, No. 22
0021-9193/05/$08.00+0     doi:10.1128/JB.187.22.7569-7578.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Unconventional Mode of Attachment of the Ruminococcus flavefaciens Cellulosome to the Cell Surface

Marco T. Rincon,1* Tadej Cepeljnik,2 Jennifer C. Martin,1 Raphael Lamed,3 Yoav Barak,4 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, Slovenia,2 Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Ramat Aviv,3 Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel4

Received 10 June 2005/ Accepted 24 August 2005

Sequence extension of the scaffoldin gene cluster from Ruminococcus flavefaciens revealed a new gene (scaE) that encodes a protein with an N-terminal cohesin domain and a C terminus with a typical gram-positive anchoring signal for sortase-mediated attachment to the bacterial cell wall. The recombinant cohesin of ScaE was recovered after expression in Escherichia coli and was shown to bind to the C-terminal domain of the cellulosomal structural protein ScaB, as well as to three unknown polypeptides derived from native cellulose-bound Ruminococcus flavefaciens protein extracts. The ScaB C terminus includes a cryptic dockerin domain that is unusual in its sequence, and considerably larger than conventional dockerins. The ScaB dockerin binds to ScaE, suggesting that this interaction occurs through a novel cohesin-dockerin pairing. The novel ScaB dockerin was expressed as a xylanase fusion protein, which was shown to bind tenaciously and selectively to a recombinant form of the ScaE cohesin. Thus, ScaE appears to play a role in anchoring the cellulosomal complex to the bacterial cell envelope via its interaction with ScaB. This sortase-mediated mechanism for covalent cell-wall anchoring of the cellulosome in R. flavefaciens differs from those reported thus far for any other cellulosome system.

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

Journal of Bacteriology, November 2005, p. 7569-7578, Vol. 187, No. 22
0021-9193/05/$08.00+0     doi:10.1128/JB.187.22.7569-7578.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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