1990 Fiscal Year Final Research Report Summary
Microbial Production and Application of Complex Carbohydrate Metabolizing Enzymes
| Project/Area Number |
01470123
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
発酵・醸造
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| Research Institution | Kyoto University |
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Principal Investigator |
TOCHIKURA Tatsurokuro Kyoto University, Department of Food Science and Technology, Professor, 農学部, 教授 (70026524)
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| 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)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Endo-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 |
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| 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.
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