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Journal of Bacteriology, September 2000, p. 4915-4925, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Scaffoldin of the Bacteroides cellulosolvens Cellulosome That Contains 11 Type II Cohesins

Shi-You Ding,1,2 Edward A. Bayer,1,* David Steiner,2 Yuval Shoham,3 and Raphael Lamed2

Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot,1 Department of Molecular Microbiology and Biotechnology, Tel-Aviv University, Ramat Aviv,2 and Department of Food Engineering and Biotechnology and Institute of Catalysis Science and Technology, Technion---Israel Institute of Technology, Haifa,3 Israel

Received 4 February 2000/Accepted 6 June 2000

A cellulosomal scaffoldin gene, termed cipBc, was identified and sequenced from the mesophilic cellulolytic anaerobe Bacteroides cellulosolvens. The gene encodes a 2,292-residue polypeptide (excluding the signal sequence) with a calculated molecular weight of 242,437. CipBc contains an N-terminal signal peptide, 11 type II cohesin domains, an internal family III cellulose-binding domain (CBD), and a C-terminal dockerin domain. Its CBD belongs to family IIIb, like that of CipV from Acetivibrio cellulolyticus but unlike the family IIIa CBDs of other clostridial scaffoldins. In contrast to all other scaffoldins thus far described, CipBc lacks a hydrophilic domain or domain X of unknown function. The singularity of CipBc, however, lies in its numerous type II cohesin domains, all of which are very similar in sequence. One of the latter cohesin domains was expressed, and the expressed protein interacted selectively with cellulosomal enzymes, one of which was identified as a family 48 glycosyl hydrolase on the basis of partial sequence alignment. By definition, the dockerins, carried by the cellulosomal enzymes of this species, would be considered to be type II. This is the first example of authentic type II cohesins that are confirmed components of a cellulosomal scaffoldin subunit rather than a cell surface anchoring component. The results attest to the emerging diversity of cellulosomes and their component sequences in nature.

* Corresponding author. Mailing address: Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel. Phone: 972-8-934-2373. Fax: 972-8-946-8256. E-mail: bfbayer{at}wicc.weizmann.ac.il.

Journal of Bacteriology, September 2000, p. 4915-4925, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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