This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Pegden, R. S. Articles by Morrison, M. Search for Related Content PubMed PubMed Citation Articles by Pegden, R. S. Articles by Morrison, M.

Journal of Bacteriology, November 1998, p. 5921-5927, Vol. 180, No. 22
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Adherence of the Gram-Positive Bacterium Ruminococcus albus to Cellulose and Identification of a Novel Form of Cellulose-Binding Protein Which Belongs to the Pil Family of Proteins

Randall S. Pegden,¹ Marilynn A. Larson,¹ Richard J. Grant,¹ and Mark Morrison^1,2,*

Department of Animal Sciences¹ and School of Biological Sciences,² University of Nebraska, Lincoln, Nebraska 68583-0908

Received 15 June 1998/Accepted 10 September 1998

The adherence of Ruminococcus albus 8 to crystalline cellulose was studied, and an affinity-based assay was also used to identify candidate cellulose-binding protein(s). Bacterial adherence in cellulose-binding assays was significantly increased by the inclusion of either ruminal fluid or micromolar concentrations of both phenylacetic and phenylpropionic acids in the growth medium, and the addition of carboxymethylcellulose (CMC) to assays decreased the adherence of the bacterium to cellulose. A cellulose-binding protein with an estimated molecular mass following sodium dodecyl sulfate-polyacrylamide gel electrophoresis of ~21 kDa, designated CbpC, was present in both cellobiose- and cellulose-grown cultures, and the relative abundance of this protein increased in response to growth on cellulose. Addition of 0.1% (wt/vol) CMC to the binding assays had an inhibitory effect on CbpC binding to cellulose, consistent with the notion that CbpC plays a role in bacterial attachment to cellulose. The nucleotide sequence of the cbpC gene was determined by a combination of reverse genetics and genomic walking procedures. The cbpC gene encodes a protein of 169 amino acids with a calculated molecular mass of 17,655 Da. The amino-terminal third of the CbpC protein possesses the motif characteristic of the Pil family of proteins, which are most commonly involved with the formation of type 4 fimbriae and other surface-associated protein complexes in gram-negative, pathogenic bacteria. The remainder of the predicted CbpC sequence was found to have significant identity with 72- and 75-amino-acid motifs tandemly repeated in the 190-kDa surface antigen protein of Rickettsia spp., as well as one of the major capsid glycoproteins of the Chlorella virus PBCV-1. Northern blot analysis showed that phenylpropionic acid and ruminal fluid increase cbpC mRNA abundance in cellobiose-grown cells. These results suggest that CbpC is a novel cellulose-binding protein that may be involved in adherence of R. albus to substrate and extends understanding of the distribution of the Pil family of proteins in gram-positive bacteria.

---

^* Corresponding author. Mailing address: C220 AnS, Marvel Baker Hall, East Campus, University of NebraskaLincoln, NE 68583-0908. Phone: (402) 472-9382. Fax: (402) 472-6362. E-mail: ansc802{at}unlvm.unl.edu.

Journal series no. 11973, Agricultural Research Division, University of Nebraska.

---

Journal of Bacteriology, November 1998, p. 5921-5927, Vol. 180, No. 22
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

• Weimer, P. J., Price, N. P. J., Kroukamp, O., Joubert, L.-M., Wolfaardt, G. M., Van Zyl, W. H. (2006). Studies of the Extracellular Glycocalyx of the Anaerobic Cellulolytic Bacterium Ruminococcus albus 7. Appl. Environ. Microbiol. 72: 7559-7566 [Abstract] [Full Text]  
• Swierczynski, A., Ton-That, H. (2006). Type III Pilus of Corynebacteria: Pilus Length Is Determined by the Level of Its Major Pilin Subunit.. J. Bacteriol. 188: 6318-6325 [Abstract] [Full Text]  
• Gaspar, A. H., Ton-That, H. (2006). Assembly of Distinct Pilus Structures on the Surface of Corynebacterium diphtheriae. J. Bacteriol. 188: 1526-1533 [Abstract] [Full Text]  
• Rakotoarivonina, H., Larson, M. A., Morrison, M., Girardeau, J.-P., Gaillard-Martinie, B., Forano, E., Mosoni, P. (2005). The Ruminococcus albus pilA1-pilA2 locus: expression and putative role of two adjacent pil genes in pilus formation and bacterial adhesion to cellulose. Microbiology 151: 1291-1299 [Abstract] [Full Text]  
• Devillard, E., Goodheart, D. B., Karnati, S. K. R., Bayer, E. A., Lamed, R., Miron, J., Nelson, K. E., Morrison, M. (2004). Ruminococcus albus 8 Mutants Defective in Cellulose Degradation Are Deficient in Two Processive Endocellulases, Cel48A and Cel9B, Both of Which Possess a Novel Modular Architecture. J. Bacteriol. 186: 136-145 [Abstract] [Full Text]  
• Koike, S., Pan, J., Kobayashi, Y., Tanaka, K. (2003). Kinetics of In Sacco Fiber-Attachment of Representative Ruminal Cellulolytic Bacteria Monitored by Competitive PCR. J DAIRY SCI 86: 1429-1435 [Abstract] [Full Text]  
• Rincon, M. T., Ding, S.-Y., McCrae, S. I., Martin, J. C., Aurilia, V., Lamed, R., Shoham, Y., Bayer, E. A., Flint, H. J. (2003). Novel Organization and Divergent Dockerin Specificities in the Cellulosome System of Ruminococcus flavefaciens. J. Bacteriol. 185: 703-713 [Abstract] [Full Text]  
• Rakotoarivonina, H., Jubelin, G., Hebraud, M., Gaillard-Martinie, B., Forano, E., Mosoni, P. (2002). Adhesion to cellulose of the Gram-positive bacterium Ruminococcus albus involves type IV pili. Microbiology 148: 1871-1880 [Abstract] [Full Text]  
• Rincon, M. T., McCrae, S. I., Kirby, J., Scott, K. P., Flint, H. J. (2001). EndB, a Multidomain Family 44 Cellulase from Ruminococcus flavefaciens 17, Binds to Cellulose via a Novel Cellulose-Binding Module and to Another R. flavefaciens Protein via a Dockerin Domain. Appl. Environ. Microbiol. 67: 4426-4431 [Abstract] [Full Text]  
• Ding, S.-Y., Rincon, M. T., Lamed, R., Martin, J. C., McCrae, S. I., Aurilia, V., Shoham, Y., Bayer, E. A., Flint, H. J. (2001). Cellulosomal Scaffoldin-Like Proteins from Ruminococcus flavefaciens. J. Bacteriol. 183: 1945-1953 [Abstract] [Full Text]