Functional Role of Aspergillus oryzae Glucoamylase C-Terminal Domain Investigated Using Its cDNA
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概要
- 論文の詳細を見る
The C-terminal domain of Aspergillus oryzae glucoamylase was analyzed by site-derected mutagenesis using glucoamylase cDNA. A mutant glucoamylase cDNA lacking the region corresponding to the C-terminal domain of the wild-type glucoamylase was constructed by inserting two stop codons in the gene for the wild-type glucoamylase. The wild-type and mutant glucoamylase cDNAs were expressed in Saccharomyces cerevisiae YPH 250,and then the produced wild-type and mutant glucomylases were purified by acarbose affinity column chromatography. As compared to those of the wild-type glucoamylase, the K_m values of the mutant enzyme determined using maltose, maltotriose or maltopentaose as a substrate were similar, but that determined using soluble starch as a substrate was twofold higher. The mutant glucoamylase showed a low rate of hydrolysis of raw cornstarch, although the wild-type glucoamylase showed a high rate of raw cornstarch hydrolysis. These results indicated that the C-terminal domain is important in the affinity of the enzyme to raw starch as reported for the glucoamylase of Aspergillus awamori.
- 社団法人日本生物工学会の論文
- 1995-09-25
著者
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Kitamoto K
Univ. Tokyo Tokyo Jpn
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Kitamoto K
National Research Institute Of Brewing
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Kitamoto Katsuhiko
Department Of Biotechnology University Of Tokyo
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Kitamoto Katsuhiko
National Research Institute Of Brewing
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KUMAGAI Chieko
National Research Institute of Brewing
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Nagashima T
Department Of Neurology Seiwa Memorial Hospital
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Nagashima Tadashi
National Research Institute Of Brewing:(present Address)technical Service Shin Nihon Chemical Co. Lt
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NAGASHIMA TADASHI
Technical service, Shin Nihon Chemical Co. Ltd.,
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YAMAMOTO YUTAKA
Technical service, Shin Nihon Chemical Co. Ltd.,
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Kumagai C
National Research Institute Of Brewing
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Kumagai Chieko
General Research Laboratories Ozeki Corp.
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Nagashima T
Teikyo Univ. School Of Medicine Ichihara Hospital
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Yamamoto Y
Division Of Integrated Life Science Graduate School Of Biostudies Kyoto University
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Yamamoto Y
Faculty Of Bio-resource Sciences Akita Prefectural University
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Kimura K
Department Of Biomedical Sciences Graduate School Of Veterinary Medicine Hokkaido University
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