This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yaoi, K.
Right arrow Articles by Mitsuishi, Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yaoi, K.
Right arrow Articles by Mitsuishi, Y.
Agricola
Right arrow Articles by Yaoi, K.
Right arrow Articles by Mitsuishi, Y.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, December 2005, p. 7670-7678, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7670-7678.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Cloning and Characterization of Two Xyloglucanases from Paenibacillus sp. Strain KM21

Katsuro Yaoi,1* Tomonori Nakai,2 Yoshiro Kameda,3 Ayako Hiyoshi,1 and Yasushi Mitsuishi1

Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan,1 Graduate School of Life Science, University of Hyogo, 2167 Shosha, Himeji, Hyogo 671-2201, Japan,2 Bioproducts Manufacturing Department, Hokkaido Sugar Co., Ltd., 3-753-2 Shinko-chuo, Ishikari, Hokkaido 061-3242, Japan3

Received 13 May 2005/ Accepted 23 July 2005

Two xyloglucan-specific endo-ß-1,4-glucanases (xyloglucanases [XEGs]), XEG5 and XEG74, with molecular masses of 40 kDa and 105 kDa, respectively, were isolated from the gram-positive bacterium Paenibacillus sp. strain KM21, which degrades tamarind seed xyloglucan. The genes encoding these XEGs were cloned and sequenced. Based on their amino acid sequences, the catalytic domains of XEG5 and XEG74 were classified in the glycoside hydrolase families 5 and 74, respectively. XEG5 is the first xyloglucanase belonging to glycoside hydrolase family 5. XEG5 lacks a carbohydrate-binding module, while XEG74 has an X2 module and a family 3 type carbohydrate-binding module at its C terminus. The two XEGs were expressed in Escherichia coli, and recombinant forms of the enzymes were purified and characterized. Both XEGs had endoglucanase active only toward xyloglucan and not toward Avicel, carboxymethylcellulose, barley ß-1,3/1,4-glucan, or xylan. XEG5 is a typical endo-type enzyme that randomly cleaves the xyloglucan main chain, while XEG74 has dual endo- and exo-mode activities or processive endo-mode activity. XEG5 digested the xyloglucan oligosaccharide XXXGXXXG to produce XXXG, whereas XEG74 digestion of XXXGXXXG resulted in XXX, XXXG, and GXXXG, suggesting that this enzyme cleaves the glycosidic bond of unbranched Glc residues. Analyses using various oligosaccharide structures revealed that unique structures of xyloglucan oligosaccharides can be prepared with XEG74.

* Corresponding author. Mailing address: Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. Phone and fax: 81-29-861-6065. E-mail: k-yaoi{at}aist.go.jp.

Applied and Environmental Microbiology, December 2005, p. 7670-7678, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7670-7678.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gloster, T. M., Ibatullin, F. M., Macauley, K., Eklof, J. M., Roberts, S., Turkenburg, J. P., Bjornvad, M. E., Jorgensen, P. L., Danielsen, S., Johansen, K. S., Borchert, T. V., Wilson, K. S., Brumer, H., Davies, G. J. (2007). Characterization and Three-dimensional Structures of Two Distinct Bacterial Xyloglucanases from Families GH5 and GH12. J. Biol. Chem. 282: 19177-19189 [Abstract] [Full Text]  
  • Martinez-Fleites, C., Guerreiro, C. I. P. D., Baumann, M. J., Taylor, E. J., Prates, J. A. M., Ferreira, L. M. A., Fontes, C. M. G. A., Brumer, H., Davies, G. J. (2006). Crystal Structures of Clostridium thermocellum Xyloglucanase, XGH74A, Reveal the Structural Basis for Xyloglucan Recognition and Degradation. J. Biol. Chem. 281: 24922-24933 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»