Applied Microbiology and Biotechnology

, Volume 53, Issue 4, pp 430–434

Cloning of the cyclodextrin glucanotransferase gene from alkalophilic Bacillus sp. A2-5a and analysis of the raw starch-binding domain

Authors

  • K. Ohdan
    • Biochemical Research Laboratory, Ezaki Glico Co., Ltd., Utajima 4-6-5, Nishiyodogawa-ku, Osaka 555-8502, Japan e-mail: kuriki-takashi@glico.co.jp Tel.: +81-6-64778425 Fax: +81-6-64778362
  • T. Kuriki
    • Biochemical Research Laboratory, Ezaki Glico Co., Ltd., Utajima 4-6-5, Nishiyodogawa-ku, Osaka 555-8502, Japan e-mail: kuriki-takashi@glico.co.jp Tel.: +81-6-64778425 Fax: +81-6-64778362
  • H. Takata
    • Biochemical Research Laboratory, Ezaki Glico Co., Ltd., Utajima 4-6-5, Nishiyodogawa-ku, Osaka 555-8502, Japan e-mail: kuriki-takashi@glico.co.jp Tel.: +81-6-64778425 Fax: +81-6-64778362
  • S. Okada
    • Biochemical Research Laboratory, Ezaki Glico Co., Ltd., Utajima 4-6-5, Nishiyodogawa-ku, Osaka 555-8502, Japan e-mail: kuriki-takashi@glico.co.jp Tel.: +81-6-64778425 Fax: +81-6-64778362
SHORT CONTRIBUTION

DOI: 10.1007/s002530051637

Cite this article as:
Ohdan, K., Kuriki, T., Takata, H. et al. Appl Microbiol Biotechnol (2000) 53: 430. doi:10.1007/s002530051637

Abstract

The cyclodextrin glucanotransferase (CGTase) gene of alkalophilic Bacillus sp. A2-5a was cloned and expressed in Bacillus subtilis ANA-1 as a host. The DNA region included an open reading frame encoding a 704-amino-acid polypeptide with a typical raw starch-binding motif in its C-terminal region. The CGTase purified from Bacillus sp. A2-5a bound to raw starch as strongly as porcine pancreas α-amylase, as expected from the sequence motif. A chromosomal region (a DNA fragment of about 14.1 kbp) including the CGTase gene was also cloned and the nucleotide sequence was determined. Possible cyclodextrinase and putative cyclodextrin-binding protein genes were found in the flanking region of the CGTase gene, which implied that the novel starch-degradation pathway postulated for a gram-negative bacterium [Klebsiella oxytoca; Fiedler et al. (1996) J Mol Biol 256: 279–291] also exists in a gram-positive bacterium i.e. Bacillus.

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© Springer-Verlag Berlin Heidelberg 2000