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 Kitamoto, N.
Right arrow Articles by Tsukagoshi, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kitamoto, N.
Right arrow Articles by Tsukagoshi, N.
Agricola
Right arrow Articles by Kitamoto, N.
Right arrow Articles by Tsukagoshi, N.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, January 1999, p. 20-24, Vol. 65, No. 1
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Sequence Analysis, Overexpression, and Antisense Inhibition of a beta -Xylosidase Gene, xylA, from Aspergillus oryzae KBN616

Noriyuki Kitamoto,1,* Shoko Yoshino,1 Kunio Ohmiya,2 and Norihiro Tsukagoshi3

Food Research Institute, Aichi Prefectural Government, Nishi-ku, Nagoya 451-0083,1 Faculty of Bioresources, Mie University, Tsu 514-8507,2 and Department of Applied Biological Sciences, Faculty of Agriculture, Nagoya University, Nagoya 464-8601,3 Japan

Received 22 June 1998/Accepted 16 October 1998

beta -Xylosidase secreted by the shoyu koji mold, Aspergillus oryzae, is the key enzyme responsible for browning of soy sauce. To investigate the role of beta -xylosidase in the brown color formation, a major beta -xylosidase, XylA, and its encoding gene were characterized. beta -Xylosidase XylA was purified to homogeneity from culture filtrates of A. oryzae KBN616. The optimum pH and temperature of the enzyme were found to be 4.0 and 60°C, respectively, and the molecular mass was estimated to be 110 kDa based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The xylA gene comprises 2,397 bp with no introns and encodes a protein consisting of 798 amino acids (86,475 Da) with 14 potential N-glycosylation sites. The deduced amino acid sequence shows high similarity to Aspergillus nidulans XlnD (70%), Aspergillus niger XlnD (64%), and Trichoderma reesei BxII (63%). The xylA gene was overexpressed under control of the strong and constitutive A. oryzae TEF1 promoter. One of the A. oryzae transformants produced approximately 13 times more of the enzyme than did the host strain. The partial-length antisense xylA gene expressed under control of the A. oryzae TEF1 promoter decreased the beta -xylosidase level in A. oryzae to about 20% of that of the host strain.

* Corresponding author. Mailing address: Food Research Institute, Aichi Prefectural Government, 2-1-1 Shinpukuji-cho, Nishi-ku, Nagoya 451-0083, Japan. Phone: 81-52-521-9316. Fax: 81-52-532-5791. E-mail: kn-afri{at}aichi-iic.or.jp.

Applied and Environmental Microbiology, January 1999, p. 20-24, Vol. 65, No. 1
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Katahira, S., Fujita, Y., Mizuike, A., Fukuda, H., Kondo, A. (2004). Construction of a Xylan-Fermenting Yeast Strain through Codisplay of Xylanolytic Enzymes on the Surface of Xylose-Utilizing Saccharomyces cerevisiae Cells. Appl. Environ. Microbiol. 70: 5407-5414 [Abstract] [Full Text]  
  • Lee, R. C., Hrmova, M., Burton, R. A., Lahnstein, J., Fincher, G. B. (2003). Bifunctional Family 3 Glycoside Hydrolases from Barley with alpha -L-Arabinofuranosidase and beta -D-Xylosidase Activity. CHARACTERIZATION, PRIMARY STRUCTURES, AND COOH-TERMINAL PROCESSING. J. Biol. Chem. 278: 5377-5387 [Abstract] [Full Text]  
  • Moralejo, F. J., Cardoza, R. E., Gutierrez, S., Lombrana, M., Fierro, F., Martin, J. F. (2002). Silencing of the Aspergillopepsin B (pepB) Gene of Aspergillus awamori by Antisense RNA Expression or Protease Removal by Gene Disruption Results in a Large Increase in Thaumatin Production. Appl. Environ. Microbiol. 68: 3550-3559 [Abstract] [Full Text]  
  • Faure, D. (2002). The Family-3 Glycoside Hydrolases: from Housekeeping Functions to Host-Microbe Interactions. Appl. Environ. Microbiol. 68: 1485-1490 [Full Text]  
  • de Vries, R. P., Visser, J. (2001). Aspergillus Enzymes Involved in Degradation of Plant Cell Wall Polysaccharides. Microbiol. Mol. Biol. Rev. 65: 497-522 [Abstract] [Full Text]  
  • La Grange, D. C., Pretorius, I. S., Claeyssens, M., van Zyl, W. H. (2001). Degradation of Xylan to D-Xylose by Recombinant Saccharomyces cerevisiae Coexpressing the Aspergillus nigerbeta -Xylosidase (xlnD) and the Trichoderma reesei Xylanase II (xyn2) Genes. Appl. Environ. Microbiol. 67: 5512-5519 [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»