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J Bacteriol, June 1998, p. 3091-3099, Vol. 180, No. 12
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Multidomain Structure and Cellulosomal Localization of the Clostridium thermocellum Cellobiohydrolase CbhA

Vladimir V. Zverlov,1 Galina V. Velikodvorskaya,1 Wolfgang H. Schwarz,2 Karin Bronnenmeier,2 Josef Kellermann,3 and Walter L. Staudenbauer2,*

Institute of Molecular Genetics, Russian Academy of Science, 123182 Moscow, Russia,1 and Institute for Microbiology, Technical University Munich, 80290 Munich,2 and Max-Planck-Institute for Biochemistry, 82152 Martinsried,3 Germany

Received 29 December 1997/Accepted 16 April 1998

The nucleotide sequence of the Clostridium thermocellum F7 cbhA gene, coding for the cellobiohydrolase CbhA, has been determined. An open reading frame encoding a protein of 1,230 amino acids was identified. Removal of a putative signal peptide yields a mature protein of 1,203 amino acids with a molecular weight of 135,139. Sequence analysis of CbhA reveals a multidomain structure of unusual complexity consisting of an N-terminal cellulose binding domain (CBD) homologous to CBD family IV, an immunoglobulin-like beta -barrel domain, a catalytic domain homologous to cellulase family E1, a duplicated domain similar to fibronectin type III (Fn3) modules, a CBD homologous to family III, a highly acidic linker region, and a C-terminal dockerin domain. The cellulosomal localization of CbhA was confirmed by Western blot analysis employing polyclonal antibodies raised against a truncated enzymatically active version of CbhA. CbhA was identified as cellulosomal subunit S3 by partial amino acid sequence analysis. Comparison of the multidomain structures indicates striking similarities between CbhA and a group of cellulases from actinomycetes. Average linkage cluster analysis suggests a coevolution of the N-terminal CBD and the catalytic domain and its spread by horizontal gene transfer among gram-positive cellulolytic bacteria.

* Corresponding author. Mailing address: Institute for Microbiology, Technical University Munich, Arcisstrasse 21, D-80290 Munich, Germany. Phone: (089) 2892-2372. Fax: (089) 2892-2360. E-mail: zverlov{at}biol.chemie.tu-muenchen.de.

J Bacteriol, June 1998, p. 3091-3099, Vol. 180, No. 12
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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