Purification and Characterization of a Protease-Resistant Cellulase from Aspergillus niger
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概要
- 論文の詳細を見る
An endo-β-1,4-glucanase (EC 3.2.1.4) was purified from a culture filtrate of Aspergillus niger IFO31125 by column chromatography through TSK-gel DEAE-3SW and TSK-gel DEAE-5PW, and by gel filtration through TSK-gel G2000SW by high performance liquid chromatography. The enzyme was estimated to have a molecular weight of about 40 kDa by both gel filtration and SDS-polyacrylamide gel electrophoresis, and appeared to consist of a monomeric protein. It contained 8.9% carbohydrate. The optimal pH for activity was 6.0-7.0,and the stable pH range was 5.0-10.0. The optimum temperature at pH 6.0 was around 70℃. The enzyme was very thermally stable and no loss of original activity was found on incubation at 60℃ for 2 h. The enzyme efficiently hydrolyzed carboxymethylcellulose and lichenan, but crystalline forms of cellulose, curdlan, laminarin, cellobiose, p-nitrophenyl-β-D-glucopyranoside and p-nitrophenyl-β-D-cellobioside were barely hydrolyzed. The activity of the enzyme was inhibited by Hg^<2+> and Cu^<2+> but was not affected by other inhibitors of thiol enzymes such as p-chloromercuribenzoate and N-ethylmaleimide. N-Bromosuccinimide showed a strong inhibitory effect, suggesting that a tryptophan residue is essential for the activity of the enzyme. The N-terminal amino acid sequence of the enzyme showed considerable homology to those of endo-β-1,4-glucanases from some other microorganisms, including Sclerotinia sclerotiorum and Schizophyllum commune. The enzyme had very strong protease-resistance, and showed no loss of activity when incubated with proteases such as Savinase at 40℃, even for 2 weeks.
- 社団法人日本生物工学会の論文
- 1995-02-25
著者
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YAMAMOTO Kenji
Department of Medical Ecology and Informatics, Research Institute, International Medical Center of J
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Yamamoto K
Graduate School Of Biostudies Kyoto University
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KUMAGAI HIDEHIKO
Department of Food Science and Technology, Kyoto University
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AKIBA SHUNICHI
Biological Science Laboratories, Kao Corporation
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Akiba Shunichi
Biological Science Laboratories Kao Corporation
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Kumagai H
Kyoto Univ.
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Kumagai H
Nihon Univ. Fujisawa‐shi Jpn
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Kumagai Hidehiko
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Kumagai Hidehiko
Faculty Of Agriculture Kyoto University
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Kumagai Hidehiko
Res. Inst. For Bioresources And Biotechnology Ishikawa Prefectural Univ.
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Kumagai Hidehiko
Division Of Integrated Life Sciences Graduate School Of Biostudies Kyoto University
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KIMURA YOSHIHARU
Biological Science Laboratories, Kao Corporation
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Kumagai H
Department Of Agricultural Chemistry The University Of Tokyo
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Kimura Yoshiharu
Biological Science Laboratories Kao Corporation
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Yamamoto Kenji
Department Of Cardiovascular Surgery Kanagawa Cardiovascular And Respiratory Center
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Yokozeki Kenzo
Research Institute For Bioresources And Biotechnology Ishikawa Prefectural University
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KUMAGAI Hidehiko
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University
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