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Cloning and sequencing of cellulase cDNA from Aspergillus kawachii and its expression in Saccharomyces cerevisiae
Authors:S. Sakamoto  G. Tamura  K. Ito  T. Ishikawa  K. Iwano  N. Nishiya
Affiliation:(1) Research Institute of Brewing Resources Co., Ltd., 2-6-30, Takinogawa, Kita-ku, 114 Tokyo, Japan;(2) National Research Institute of Brewing, 2-6-30, Takinogawa, Kita-ku, 114 Tokyo, Japan;(3) Present address: Meiwa Shoji Co., Ltd., 1-14-2 Shinjuku, Shinjuku-ku, 160 Tokyo, Japan
Abstract:
The cDNA encoding the endo-beta-1,4-glucanase (carboxymethylcellulase; CMCase-I) from Aspergillus kawachii IFO 4308 was cloned. Nucleotide-sequence analysis of the cloned cDNA insert showed a 717-bp open reading frame that encoded a protein of 239 amino-acid residues. The predicted amino-acid sequence of the mature protein had considerable homology with the protein sequence of the FI-CMCase of Aspergillus aculeatus. The cDNA was introduced into Saccharomyces cerevisiae. The expressed enzyme had carboxylmethylcellulase acitivity, identified by clear zones on a CMC-agar plate after Congo Red staining.
Keywords:Cellulase  Aspergillus kawachii  cDNA cloning  Expression
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