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1990 Fiscal Year Final Research Report Summary

Microbial Production and Application of Complex Carbohydrate Metabolizing Enzymes

Research Project

Project/Area Number 01470123
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 発酵・醸造
Research InstitutionKyoto University

Principal Investigator

TOCHIKURA Tatsurokuro  Kyoto University, Department of Food Science and Technology, Professor, 農学部, 教授 (70026524)

Co-Investigator(Kenkyū-buntansha) YAMAMOTO Kenji  Kyoto Univ., Dept. of Food Sci. & Technol., Research Associate, 農学部, 助手 (70109049)
SUZUKI Hideyuki  Kyoto Univ., Dept. of Food Sci. & Technol., Research Associate, 農学部, 助手 (10202136)
KUMAGAI Hidehiko  Kyoto Univ., Dept. of Food Sci. & Technol., Associate Professor, 農学部, 助教授 (70027192)
YANO Toshihiro  Kyoto Univ., Dept. of Food Sci. & Technol., Research Official, 農学部, 教務職員 (30135553)
Project Period (FY) 1989 – 1990
KeywordsEndo-beta-N-acetylglucosaminidase / Complex-type sugar chain / A-type blood substance / alpha-N-acetylgalactosaminidase / alpha-L-Fucosidase / Endo-alpha-N-acetylgalactosaminidase / Human gastric carcinoma cell / T-antigen
Research Abstract

1. A novel endo-beta-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. The enzyme, named Endo-M, was purified to almost homogeneity by polyacrylamide gel electrophoresis.
2. A fungus isolated from soil was found to produce alpha-N-acetylgalactosaminidase in culture, this enzyme being the dominant glycosidase. The fungus was identified as an Acremonium sp. The enzyme produced by the fungus was purified to electrophoretic homogeneity from the culture broth. The enzyme could change blood type A erythrocytes into blood type O erythrocytes.
3. Bacillus circulans isolated from soil was found to produce two types of alpha-L-fucosidase differing in substrate specificity. One was able to liberate L-fucose from Porcine Gastric Mucin (PGM), but not from artificial substrates, including p-nitrophenyl and methyl alpha-L-fucosides, while the other acted not on PGM but on p-nitrophenyl alpha-L-fucoside. The production of the former enzyme was enhanced about 150 times as much by PGM added to the medium as by glucose.
4. Two forms of alpha-L-fucosidase, deglycosylated and glycosylated, were found in the fucose-inducing culture broth of Fusarium oxysporum. Endo-beta-N-acetylglucosaminidase was also found in the same culture broth. Results suggest that the deglycosylated alpha-L-fucosidase is formed by the release of sugar chains from the glycosylated form by Fusarium endo-beta-N-acetylglucosaminidase.
5. (1) Endo-alpha-N-acetylgalactosaminidase from Alcaligenes sp. released the disaccharide, Galbeta1->3GalNAc (T-antigen), from both dansylated serine-GalNAc-Gal and threonine-GalNAc-Gal, and showed higher activity on the former than the latter.
(2) The released T-antigen from human gastric carcinoma cell Kato III could be analyzed quantitatively by a sensitive method including pyridylamino derivatization and following high performance liquid chromatography.

  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] S.Kadowaki: "Isolation and Characterization of a Blood Group A SubstanceーDegrading αーNーAcetylgalactosaminidase from an Acremonium sp." Agricultural and Biological Chemistry. 53. 111-120 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Takegawa: "Primary Structure of an NーLinked Sugar Chain Derived from Glucoamylase of Rhizopus niveus." Biochimica et Biphysica Acta. 990. 98-100 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Kadowaki: "Purification and Characterization of a Novel Fungal EndoーβーNーacetylglucosaminidase Acting on Complex Oligosaccharides of Glycoproteins." Agricultural and Biological Chemistry. 54. 97-106 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.ーQ.Fan: "Induction and Efficient Purification of EndoーαーNーacetylgalactosaminidase from Alcaligenes sp." Agricultural and Biological Chemistry. 54. 233-234 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Tsuji: "Purification and Characterization of αーLーFucosidase from Bacillus circulans Grown on Porcine Gastric Mucin." Journal of Biochemistry. 107. 324-330 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Tsuji: "Formation of Deglycosylated αーLーFucosidase by EndoーβーNーacetylglucosaminidase in Fusarium oxysporum." Applied and Environmental Microbiology. 56. 928-933 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.ーQ.Fan: "Action of EndoーαーNーacetylgalactosaminidase from Alcaligenes sp.on AminoーacidーOーglucans;Comparison with the Enzyme from Diplococcus pneumoniae." Biochemcal and Biophysical Research Communications. 169. 751-757 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Tsuji: "Purification and Some Properties of a Novel αーLーFucosidase Capable of Acting on αー(1→6)ーLーFucosidic Linkages from Bacillus circulans M28." Journal of Biochemistry. 108. 235-240 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.ーQ.Fan: "Release of Tーantigen, a Carcinoma Marker from Native Human Cells,by EndoーαーNーacetylgalactosaminidase of Alcaligenes sp." Biochemical and Biophysical Research Communications. 172. 341-347 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S. Kadowaki, T. Ueda, K. Yamamoto, H. Kumagai and T. Tochikura.: "Isolation and characterization of a blood group A substance-degrading alpha-N-acetylgalactosaminidase from an Acremonium sp." Agricultural and Biochemical Chemistry. 53. 111-120 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Takegawa, N. Kawasaki, S. Iwahara, K. Yamamoto, T. Tochikura, B. Mikami and Y. Morita.: "Primary structure of an N-linked sugar chain derived from glucoamylase of Rhizopus niveus." Biochimca et Biophysica Acta. 990. 98-100 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Kadowaki, K. Yamamoto, M. Fijisaki, K. Izumi, T. Tochikura and T. Yokoyama.: "Purification and characterization of a novel fungal endo-beta-N-acetylglucosaminidase acting on complex oligosaccharides of glycoproteins." Agricultural and Biochemical Chemistry. 54. 97-106 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J. -Q. Fan, K. Yamamoto, H. Kumagai and T. Tochikura.: "Induction and efficient purification of endo-alpha-N-acetylgalactosaminidase from Alcaligenes sp." Agricultural and Biochemical Chemistry. 54. 233-234 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y. Tsuji, K. Yamamoto, T. Tochikura, T. Senoo, Y. Ohkubo and H. Yamaguchi.: "Purification and characterization of alpha-L-fucosidase from Bacillus circulants grown on porcine gastric mucin." Journal of Biochemistry. 107. 324-330 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y. Tsuji, K. Yamamoto and T. Tochikura.: "Formation of deglycosylated alpha-L-fucosidase by endo-beta-N-acetyglucosaminidase in Fusarium oxysporum." Applied and Environmental Microbiology. 56. 928-933 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J. -Q. Fan, K. Yamamoto, Y. Matsumoto, Y. Hirabayashi H. Kumagai and T. Tochikura.: "Action of endo-alpha-N-acetylgalactosaminidase from Alcaligenes sp. on amino acide-O-glucans : comparison with the enzyme from Diprococcus pneumoniae." Biochemical and Biophysical Research Communications. 169. 751-757 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y. Tsuji, K. Yamamoto and T. Tochikura.: "Purification and some properties of a novel alpha-L-fucosidase capable of action on alpha-(1->6)-L-fucosidic linkages from Bacillus circulants M28." Journal of Biochemistry. 108. 235-240 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J. -Q. Fan, K. Kanaya, N. Ando, K. Yamamoto, H. Kumagai and T. Tochikura.: "Release of T-antigen, a carcinoma maker from native human cells, by endo-alpha-N-acetylgalactosaminidase of Alcaligenes sp." Biochemical and Biophysical Research Communications. 172. 341-347 (1990)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-08-12  

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