Purification and Characterization of a Novel Fungal Endo-β-N-acetylglucosaminidase Acting on Complex Oligosaccharides of Glycoproteins(Microbiology & Fermentation Industry)
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概要
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A novel endo-β-N-acetylglucosaminidase acting on complex type sugar chains of glycoproteins was found in the culture broth of a fungus isolated from soil and identified as Mucor hiemalis f. hiemalis on the basis of various characteristics. The endo-β-N-acetylglucosaminidase, named Endo-M, was purified to almost homogeneity by polyacrylamide gel electrophoresis involving ammonium sulfate fractionation, and column chromatographies on DEAE-Sepharose CL-6B, Sephadex G-200, hydroxylapatite, TSK-gel HW-65F and Con A-Sepharose 4B. The molecular weight of the enzyme was estimated to be about 95,000 by gel chromatography. The optimum pH was found to be 6.0-6.5 and the enzyme was stable in the pH range of 7 to 8. The enzyme showed high activity on dansyl ovalbumin glycopeptide, and also could act on dansyl transferrin glycopeptide and dansyl asialotransferrin glycopeptide containing biantennary complex type sugar chains. The Km value for dansyl asialotransferrin glycopeptide as the substrate was 2.0×10^<-3> M. The enzyme released complex type sugar chains from intact asialotransferrin without the addition of any detergent and the liberated sugar chains were identified by chromatography on a Bio-Gel P-4 column, calibrated with markers of known structure, and ^1H-NMR analysis.
- 社団法人日本農芸化学会の論文
- 1990-01-23
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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TOCHIKURA TATSUROKURO
Department of Food Science and Technology, Kyoto University
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KADOWAKI Setsu
Kacho College
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FUJISAKI Masatoki
Department of Food Science and Technology, Kyoto University
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Yokoyama Tatsuo
Institute For Fermentation Osaka
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Izumi Kazuo
Government Industrial Development Laboratory
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Tochikura Tatsurokuro
Faculty Of Home Economics Kobe Women's University
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Tochikura Tatsurokuro
Department Of Food Science And Technology Kyoto Unicersity
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Tochikura Tatsurokuro
Department Of Agricultural Chemistry Kyoto University
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Kadowaki S
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University Kacho Junior Colle
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Tachiki Takashi
Department Of Chemistry Ritsumeikan University
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Yano Toshihiro
Department Of Food Science Ishikawa Agricultural College
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Tachiki T
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Tachiki Takashi
Faculty Of Home Economics Kobe Women's University
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Tachiki Tkashi
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Yano T
Research Laboratory Of Higashimaru Shoyu Co. Ltd.
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Fujisaki Masatoki
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Yamamoto Kenji
Department Of Cardiovascular Surgery Kanagawa Cardiovascular And Respiratory Center
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