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 Hashimoto, M.
Right arrow Articles by Watanabe, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hashimoto, M.
Right arrow Articles by Watanabe, T.

 Previous Article  |  Next Article 

Journal of Bacteriology, June 2000, p. 3045-3054, Vol. 182, No. 11
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Expression and Characterization of the Chitin-Binding Domain of Chitinase A1 from Bacillus circulans WL-12

Masayuki Hashimoto,1 Takahisa Ikegami,2 Shizuka Seino,1 Nobuhumi Ohuchi,3 Harumi Fukada,4 Junji Sugiyama,5 Masahiro Shirakawa,2 and Takeshi Watanabe1,3,*

Department of Biosystem Science, Graduate School of Science and Technology,1 and Department of Applied Biological Chemistry, Faculty of Agriculture,3 Niigata University, 8050 Ikarashi-2, Niigata 950-2181, Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101,2 Laboratory of Biophysical Chemistry, College of Agriculture, Osaka Prefecture University, Sakai, Osaka 599-8531,4 and Wood Research Institute, Kyoto University, Uji, Kyoto, 611-0011,5 Japan

Received 12 July 1999/Accepted 5 March 2000

Chitinase A1 from Bacillus circulans WL-12 comprises an N-terminal catalytic domain, two fibronectin type III-like domains, and a C-terminal chitin-binding domain (ChBD). In order to study the biochemical properties and structure of the ChBD, ChBDChiA1 was produced in Escherichia coli using a pET expression system and purified by chitin affinity column chromatography. Purified ChBDChiA1 specifically bound to various forms of insoluble chitin but not to other polysaccharides, including chitosan, cellulose, and starch. Interaction of soluble chitinous substrates with ChBDChiA1 was not detected by means of nuclear magnetic resonance and isothermal titration calorimetry. In addition, the presence of soluble substrates did not interfere with the binding of ChBDChiA1 to regenerated chitin. These observations suggest that ChBDChiA1 recognizes a structure which is present in insoluble or crystalline chitin but not in chito-oligosaccharides or in soluble derivatives of chitin. ChBDChiA1 exhibited binding activity over a wide range of pHs, and the binding activity was enhanced at pHs near its pI and by the presence of NaCl, suggesting that the binding of ChBDChiA1 is mediated mainly by hydrophobic interactions. Hydrolysis of beta -chitin microcrystals by intact chitinase A1 and by a deletion derivative lacking the ChBD suggested that the ChBD is not absolutely required for hydrolysis of beta -chitin microcrystals but greatly enhances the efficiency of degradation.

* Corresponding author. Mailing address: Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, 8050 Ikarashi-2, Niigata 950-2181, Japan. Phone: (81)252626647. Fax: (81)252626854. E-mail: wata{at}agr.niigata-u.ac.jp.

Journal of Bacteriology, June 2000, p. 3045-3054, Vol. 182, No. 11
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Akagi, K.-i., Watanabe, J., Hara, M., Kezuka, Y., Chikaishi, E., Yamaguchi, T., Akutsu, H., Nonaka, T., Watanabe, T., Ikegami, T. (2006). Identification of the Substrate Interaction Region of the Chitin-Binding Domain of Streptomyces griseus Chitinase C.. J Biochem 139: 483-493 [Abstract] [Full Text]  
  • Wang, J.-Y., Chao, Y.-P. (2006). Immobilization of Cells with Surface-Displayed Chitin-Binding Domain. Appl. Environ. Microbiol. 72: 927-931 [Abstract] [Full Text]  
  • Colussi, P. A., Specht, C. A., Taron, C. H. (2005). Characterization of a Nucleus-Encoded Chitinase from the Yeast Kluyveromyces lactis. Appl. Environ. Microbiol. 71: 2862-2869 [Abstract] [Full Text]  
  • Vaaje-Kolstad, G., Houston, D. R., Riemen, A. H. K., Eijsink, V. G. H., van Aalten, D. M. F. (2005). Crystal Structure and Binding Properties of the Serratia marcescens Chitin-binding Protein CBP21. J. Biol. Chem. 280: 11313-11319 [Abstract] [Full Text]  
  • Kojima, M., Yoshikawa, T., Ueda, M., Nonomura, T., Matsuda, Y., Toyoda, H., Miyatake, K., Arai, M., Fukamizo, T. (2005). Family 19 Chitinase from Aeromonas sp. No.10S-24: Role of Chitin-Binding Domain in the Enzymatic Activity. J Biochem 137: 235-242 [Abstract] [Full Text]  
  • Katouno, F., Taguchi, M., Sakurai, K., Uchiyama, T., Nikaidou, N., Nonaka, T., Sugiyama, J., Watanabe, T. (2004). Importance of Exposed Aromatic Residues in Chitinase B from Serratia marcescens 2170 for Crystalline Chitin Hydrolysis. J Biochem 136: 163-168 [Abstract] [Full Text]  
  • Kobayashi, D. Y., Reedy, R. M., Bick, J., Oudemans, P. V. (2002). Characterization of a Chitinase Gene from Stenotrophomonas maltophilia Strain 34S1 and Its Involvement in Biological Control. Appl. Environ. Microbiol. 68: 1047-1054 [Abstract] [Full Text]  
  • Wu, M. L., Chuang, Y. C., Chen, J. P., Chen, C. S., Chang, M. C. (2001). Identification and Characterization of the Three Chitin-Binding Domains within the Multidomain Chitinase Chi92 from Aeromonas hydrophila JP101. Appl. Environ. Microbiol. 67: 5100-5106 [Abstract] [Full Text]  
  • Thompson, S. E., Smith, M., Wilkinson, M. C., Peek, K. (2001). Identification and Characterization of a Chitinase Antigen from Pseudomonas aeruginosa Strain 385. Appl. Environ. Microbiol. 67: 4001-4008 [Abstract] [Full Text]  
  • Saito, A., Miyashita, K., Schrempf, H. (2001). Characteristics of a Streptomyces coelicolor A3(2) Extracellular Protein Targeting Chitin and Chitosan. Appl. Environ. Microbiol. 67: 1268-1273 [Abstract] [Full Text]  
  • Uchiyama, T., Katouno, F., Nikaidou, N., Nonaka, T., Sugiyama, J., Watanabe, T. (2001). Roles of the Exposed Aromatic Residues in Crystalline Chitin Hydrolysis by Chitinase A from Serratia marcescens 2170. J. Biol. Chem. 276: 41343-41349 [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»