Applied Microbiology and Biotechnology

, Volume 39, Issue 2, pp 197–203

Cloning and sequence analysis of the cyclomaltodextrinase gene from Bacillus sphaericus and expression in Escherichia coli cells

  • Tetsuya Oguma
  • Asahi Matsuyama
  • Mamoru Kikuchi
  • Eiichi Nakano
Article

Abstract

The gene for cyclomaltodextrinase (CDase; EC 3.2.1.54) from Bacillus sphaericus E-244 was cloned in the recombinant plasmid pCD629. Sequencing a portion of pCD629 revealed a unique open reading frame of 1,773 nucleotides coding for a 591-amino-acid polypeptide. The deduced polypeptide sequence showed about 50% homology with that of a neopullulanase, and was slightly homologous to those of the cyclodextrin glucanotransferases and the α-amylases. The optimum pH, specific activity and Km value for β-cyclodextrin of the CDase that has been produced in Escherichia coli cells were 8.0, 16.4 units/mg protein, and 0.41 mm, respectively. These values were almost identical to those from B. sphaericus E-244.

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Copyright information

© springer-Verlag 1993

Authors and Affiliations

  • Tetsuya Oguma
    • 1
  • Asahi Matsuyama
    • 1
  • Mamoru Kikuchi
    • 2
  • Eiichi Nakano
    • 2
  1. 1.Noda Institute for Scientific ResearchNoda City, Chiba Pref.Japan
  2. 2.Research and Development DivisionKikkoman CorporationNoda City, Chiba Pref.Japan

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