Cloning and sequence analysis of the cyclomaltodextrinase gene from Bacillus sphaericus and expression in Escherichia coli cells |
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| Authors: | Tetsuya Oguma Asahi Matsuyama Mamoru Kikuchi Eiichi Nakano |
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| Institution: | (1) Noda Institute for Scientific Research, 399 Noda, 278 Noda City, Chiba Pref., Japan;(2) Research and Development Division, Kikkoman Corporation, 399 Noda, 278 Noda City, Chiba Pref., Japan |
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| 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  -amylases. The optimum pH, specific activity and K
m value for  -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.
Correspondence to: T. Oguma |
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