Cloning and sequencing of cellulase cDNA from Aspergillus kawachii and its expression in Saccharomyces cerevisiae.
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 activity, identified by clear zones on a CMC-agar plate after Congo Red staining.
Sakamoto
S
S
Research Institute of Brewing Resources Co., Ltd., Tokyo, Japan.
Tamura
G
G
Ito
K
K
Ishikawa
T
T
Iwano
K
K
Nishiya
N
N
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002480:Cellulase / Q000737:chemistry* / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic
D018076:DNA, Complementary; D002480:Cellulase