1998 Fiscal Year Annual Research Report

エンド型グリコシダーゼの特異的な活性を利用した新しい糖鎖工学的研究の展開

Research Project

Project/Area Number	09358013
Research Category	Grant-in-Aid for Scientific Research (A)
Research Institution	Hirosaki University
Principal Investigator	遠藤正彦弘前大学, 医学部, 教授 (20004616)
Co-Investigator(Kenkyū-buntansha)	竹川薫香川大学, 農学部, 助教授 (50197282) 伊東信九州大学, 農学部, 助教授 (40253512) 山本憲二京都大学, 大学院・農学研究科, 助教授 (70109049) 長谷純宏大阪大学, 大学院・理学研究科, 教授 (80028232) 堀田恭子北里大学, 医学部, 教授 (10050402)
Keywords	エンド型グリコシダーゼ / 糖鎖工学 / 糖タンパク質 / 糖脂質 / プロテオグリカン / 糖鎖切り出し / 糖鎖組み換え / 糖鎖限定分解
Research Abstract	1. 研究の背景と目的:本研究組織の構成員は、それぞれ自らが、エンド型グリコシダーゼの発見者である。その酵素は、複合糖質のタンパク質(または脂質)と糖鎖との結合部位を選択的に分解するという特異性を有する。したがって、本酵素を糖鎖の限定分解や糖の転移に利用すると、新しい糖鎖工学的研究展開の道が拓かれる。本研究は、その確立を目的としている。 2. 研究の成果:研究の成果について、公開シンポジウムを開催して報告している。 1) エンド-β-キシロシダーゼは、プロテオグリカンのペプチドとグリコサミノグリカンの結合部位に作用する。本酵素を用いた糖鎖の切り出し法とその糖鎖工学的応用法が検討されている(遠藤、高垣)。 2) エンド-α-N-アセチルガラクトサミニダーゼは、ムチン型糖タンパク質のペプチドと糖鎖の結合部位に作用する。本酵素を産生する細菌の培養条件、及び酵素の分解法が検討されている(堀田)。 3) エンド-β-N-アセチルグルコサミニダーゼは、N-グリコシド型糖タンパク質のペプチドと糖鎖の結合部位に作用する。この酵素群の中でエンド-M酵素(山本)、エンド-A酵素(竹川)及びエンド-HS酵素(伊藤)を用いて糖転移を行い新規糖ペプチドが合成された。 4) エンド-β-マンノシダーゼは、N-グリコシド型糖鎖のマンノシド結合を切る酵素として新しく発見された(長谷)。 5) エンドグリコセラミダーゼは、脂質と糖鎖の結合部位を切る遺伝子がクローニングされた(伊東)。 3. 今後の研究展開:今後、クローニングと糖鎖工学的な応用展開に向けて研究が進められる。

Research Products
(16 results)

All Other

All Publications (16 results)

[Publications] Takagaki,K.: "Ion-spray mass spectromentry for identification of the nonreducing terminal sugar of glycosaminoglycan." Glycobiology. 8. 719-724 (1998)
[Publications] Takagaki,K.: "Characterization of β-D-xyloside-initiated glycosaminoglycan synthesized by human skin fibroblasts in the presence of tunicamycin." Glycoconjugate J.15. 483-489 (1998)
[Publications] Tazawa,T.: "A novel 4-methylumbelliferyi-β-D-xyloside derivative,sulfate-O-3-xylosyl β1-(4-methylumbelliferone),isolated from culture medium of human skin fihroblasts, and its role in 4-methylumbelliferone-initiated glycosaminoglycan biosynthesis." Glycobiology. 8. 879-884 (1998)
[Publications] Ishii-Karakasa,I.: "H-NMR analysis of 2-pyridylaminated O-linked sugar chains from fetuin in fetal calf serum." Bunseki Kagaku. 46. 101-110 (1997)
[Publications] Tanaka,T.: "Screening and Fermentation of Endo-α-N-Acetylgalactosaminidase S,a Mucin-Hydrolyzing Enzyme from Streptomyces Acting on the GalNAc-O-Ser(Thr)Linkage." Journal of Fermentation and Bioengineering. 85. 381-387 (1998)
[Publications] Iwase,H.: "Application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry to the analysis of glycopeptide-containing multiple O-linked oligosaccharides." Journal of Chromatography B.709. 145-149 (1998)
[Publications] Sasaki,A.: "Partial Purification and Characterization of a Novel Endo-β-mannosidase Acting on N-linked Sugar Chains from Lilium longflorum Thumb." J.Biochem. in press. 125. (1999)
[Publications] Iwai,K.: "Detection of Man_5GlcNAc and Related Free Oligosaccharides in the Cytosol Fraction of Hen Oviduct." J.Biochem.125. 70-74 (1999)
[Publications] Yamamoto,K.: "Chemo-enzymatic Synthesis of a Novel Glycopeptide Using a Microbial Endoglycosidase." Carbohyd.Res.305. 415-422 (1998)
[Publications] Haneda,K.: "Chemo-enzymatic Synthesis of Calcitonin Derivatives Containing N-Linked Oligosaccharides." Bioorg.Med.Chem.Lett.8. 1303-1306 (1998)
[Publications] Okino,N.: "Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa." J.Biol Chem.273. 14368-14373 (1998)
[Publications] Komori,H.: "Regulation of intracellular ceramide content in B16 melanoma cells-BIOLOGICAL IMPLICATIONS OF CERAMIDE GLYCOSYLATION-" J.Biol.Chem.274(in press)(April issue).
[Publications] Takegawa,K.: "Enzymatic synthesis of a neoglycoconjugate by transglycosylation with Arthrobacter endo-β-N-acetylglucosaminidase ; a substrate for colorimetric detection of endo-β-N-acetylglucosaminidase activity." Anal.Biochem.257. 218-223 (1998)
[Publications] Deras,I.L.: "Synthesis of a high-mannose-type oligosaccharide glycopeptide analog containing a glucose-asparagine linkage." Bioorg.Med.Chem.Lett.8. 1763-1766 (1998)
[Publications] Renu,V.: "Evaluation of Enzyme-Linked Immunosorbent Assay for the Detection of Cymbisium Mosaic Virus in Orchids." J.Ferment.Bioeng.86. 65-71 (1998)
[Publications] Trisanti,A.: "Three Different Types of α-Amylases from Aspergillus awamori KT-11 ; Their Purifications,Properties,and Specificities." Biosci.Biotechnol.Biochem.62. 1351-1357 (1998)

1998 Fiscal Year Annual Research Report

エンド型グリコシダーゼの特異的な活性を利用した新しい糖鎖工学的研究の展開

Principal Investigator

遠藤 正彦 弘前大学, 医学部, 教授 (20004616)

Research Products

[Publications] Takagaki,K.: "Ion-spray mass spectromentry for identification of the nonreducing terminal sugar of glycosaminoglycan." Glycobiology. 8. 719-724 (1998)

[Publications] Takagaki,K.: "Characterization of β-D-xyloside-initiated glycosaminoglycan synthesized by human skin fibroblasts in the presence of tunicamycin." Glycoconjugate J.15. 483-489 (1998)

[Publications] Tazawa,T.: "A novel 4-methylumbelliferyi-β-D-xyloside derivative,sulfate-O-3-xylosyl β1-(4-methylumbelliferone),isolated from culture medium of human skin fihroblasts, and its role in 4-methylumbelliferone-initiated glycosaminoglycan biosynthesis." Glycobiology. 8. 879-884 (1998)

[Publications] Ishii-Karakasa,I.: "H-NMR analysis of 2-pyridylaminated O-linked sugar chains from fetuin in fetal calf serum." Bunseki Kagaku. 46. 101-110 (1997)

[Publications] Tanaka,T.: "Screening and Fermentation of Endo-α-N-Acetylgalactosaminidase S,a Mucin-Hydrolyzing Enzyme from Streptomyces Acting on the GalNAc-O-Ser(Thr)Linkage." Journal of Fermentation and Bioengineering. 85. 381-387 (1998)

[Publications] Iwase,H.: "Application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry to the analysis of glycopeptide-containing multiple O-linked oligosaccharides." Journal of Chromatography B.709. 145-149 (1998)

[Publications] Sasaki,A.: "Partial Purification and Characterization of a Novel Endo-β-mannosidase Acting on N-linked Sugar Chains from Lilium longflorum Thumb." J.Biochem. in press. 125. (1999)

[Publications] Iwai,K.: "Detection of Man_5GlcNAc and Related Free Oligosaccharides in the Cytosol Fraction of Hen Oviduct." J.Biochem.125. 70-74 (1999)

[Publications] Yamamoto,K.: "Chemo-enzymatic Synthesis of a Novel Glycopeptide Using a Microbial Endoglycosidase." Carbohyd.Res.305. 415-422 (1998)

[Publications] Haneda,K.: "Chemo-enzymatic Synthesis of Calcitonin Derivatives Containing N-Linked Oligosaccharides." Bioorg.Med.Chem.Lett.8. 1303-1306 (1998)

[Publications] Okino,N.: "Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa." J.Biol Chem.273. 14368-14373 (1998)

[Publications] Komori,H.: "Regulation of intracellular ceramide content in B16 melanoma cells-BIOLOGICAL IMPLICATIONS OF CERAMIDE GLYCOSYLATION-" J.Biol.Chem.274(in press)(April issue).

[Publications] Takegawa,K.: "Enzymatic synthesis of a neoglycoconjugate by transglycosylation with Arthrobacter endo-β-N-acetylglucosaminidase ; a substrate for colorimetric detection of endo-β-N-acetylglucosaminidase activity." Anal.Biochem.257. 218-223 (1998)

[Publications] Deras,I.L.: "Synthesis of a high-mannose-type oligosaccharide glycopeptide analog containing a glucose-asparagine linkage." Bioorg.Med.Chem.Lett.8. 1763-1766 (1998)

[Publications] Renu,V.: "Evaluation of Enzyme-Linked Immunosorbent Assay for the Detection of Cymbisium Mosaic Virus in Orchids." J.Ferment.Bioeng.86. 65-71 (1998)

[Publications] Trisanti,A.: "Three Different Types of α-Amylases from Aspergillus awamori KT-11 ; Their Purifications,Properties,and Specificities." Biosci.Biotechnol.Biochem.62. 1351-1357 (1998)

遠藤正彦弘前大学, 医学部, 教授 (20004616)