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 Terada, Y.
Right arrow Articles by Okada, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Terada, Y.
Right arrow Articles by Okada, S.
Agricola
Right arrow Articles by Terada, Y.
Right arrow Articles by Okada, S.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, March 1999, p. 910-915, Vol. 65, No. 3
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Thermus aquaticus ATCC 33923 Amylomaltase Gene Cloning and Expression and Enzyme Characterization: Production of Cycloamylose

Yoshinobu Terada,* Kazutoshi Fujii, Takeshi Takaha, and Shigetaka Okada

Biochemical Research Laboratory, Ezaki Glico Co., Ltd., 4-6-5 Utajima, Nishiyodogawa, Osaka 555-8502, Japan

Received 6 August 1998/Accepted 1 December 1998

The amylomaltase gene of the thermophilic bacterium Thermus aquaticus ATCC 33923 was cloned and sequenced. The open reading frame of this gene consisted of 1,503 nucleotides and encoded a polypeptide that was 500 amino acids long and had a calculated molecular mass of 57,221 Da. The deduced amino acid sequence of the amylomaltase exhibited a high level of homology with the amino acid sequence of potato disproportionating enzyme (D-enzyme) (41%) but a low level of homology with the amino acid sequence of the Escherichia coli amylomaltase (19%). The amylomaltase gene was overexpressed in E. coli, and the enzyme was purified. This enzyme exhibited maximum activity at 75°C in a 10-min reaction with maltotriose and was stable at temperatures up to 85°C. When the enzyme acted on amylose, it catalyzed an intramolecular transglycosylation (cyclization) reaction which produced cyclic alpha -1,4-glucan (cycloamylose), like potato D-enzyme. The yield of cycloamylose produced from synthetic amylose with an average molecular mass of 110 kDa was 84%. However, the minimum degree of polymerization (DP) of the cycloamylose produced by T. aquaticus enzyme was 22, whereas the minimum DP of the cycloamylose produced by potato D-enzyme was 17. The T. aquaticus enzyme also catalyzed intermolecular transglycosylation of maltooligosaccharides. A detailed analysis of the activity of T. aquaticus ATCC 33923 amylomaltase with maltooligosaccharides indicated that the catalytic properties of this enzyme differ from those of E. coli amylomaltase and the plant D-enzyme.

* Corresponding author. Mailing address: Biochemical Research Laboratory, Ezaki Glico Co., Ltd., 4-6-5 Utajima, Nishiyodogawa, Osaka 555-8502, Japan. Phone: 81-6-6477-8425. Fax: 81-6-6477-8271. E-mail: terada-yoshinobu{at}glico.co.jp.

Applied and Environmental Microbiology, March 1999, p. 910-915, Vol. 65, No. 3
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Barends, T. R. M., Bultema, J. B., Kaper, T., van der Maarel, M. J. E. C., Dijkhuizen, L., Dijkstra, B. W. (2007). Three-way Stabilization of the Covalent Intermediate in Amylomaltase, an {alpha}-Amylase-like Transglycosylase. J. Biol. Chem. 282: 17242-17249 [Abstract] [Full Text]  
  • Fujii, K., Minagawa, H., Terada, Y., Takaha, T., Kuriki, T., Shimada, J., Kaneko, H. (2005). Use of Random and Saturation Mutageneses To Improve the Properties of Thermus aquaticus Amylomaltase for Efficient Production of Cycloamyloses. Appl. Environ. Microbiol. 71: 5823-5827 [Abstract] [Full Text]  
  • Kaper, T., Talik, B., Ettema, T. J., Bos, H., van der Maarel, M. J. E. C., Dijkhuizen, L. (2005). Amylomaltase of Pyrobaculum aerophilum IM2 Produces Thermoreversible Starch Gels. Appl. Environ. Microbiol. 71: 5098-5106 [Abstract] [Full Text]  
  • Imamura, H., Fushinobu, S., Yamamoto, M., Kumasaka, T., Jeon, B.-S., Wakagi, T., Matsuzawa, H. (2003). Crystal Structures of 4-{alpha}-Glucanotransferase from Thermococcus litoralis and Its Complex with an Inhibitor. J. Biol. Chem. 278: 19378-19386 [Abstract] [Full Text]  
  • Yanase, M., Takata, H., Takaha, T., Kuriki, T., Smith, S. M., Okada, S. (2002). Cyclization Reaction Catalyzed by Glycogen Debranching Enzyme (EC 2.4.1.25/EC 3.2.1.33) and Its Potential for Cycloamylose Production. Appl. Environ. Microbiol. 68: 4233-4239 [Abstract] [Full Text]  
  • Kamasaka, H., Sugimoto, K., Takata, H., Nishimura, T., Kuriki, T. (2002). Bacillus stearothermophilus Neopullulanase Selective Hydrolysis of Amylose to Maltose in the Presence of Amylopectin. Appl. Environ. Microbiol. 68: 1658-1664 [Abstract] [Full Text]  
  • Terada, Y., Sanbe, H., Takaha, T., Kitahata, S., Koizumi, K., Okada, S. (2001). Comparative Study of the Cyclization Reactions of Three Bacterial Cyclomaltodextrin Glucanotransferases. Appl. Environ. Microbiol. 67: 1453-1460 [Abstract] [Full Text]  
  • Duffner, F., Bertoldo, C., Andersen, J. T., Wagner, K., Antranikian, G. (2000). A New Thermoactive Pullulanase from Desulfurococcus mucosus: Cloning, Sequencing, Purification, and Characterization of the Recombinant Enzyme after Expression in Bacillus subtilis. J. Bacteriol. 182: 6331-6338 [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»