Glycotechnological Studies on Oligosaccharide Units Involved in Molecular Recognition.

参与分子识别的寡糖单元的糖技术研究。

• 批准号: 06808055
• 负责人: USUI Taichi
• 金额: $ 1.28万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06808055/
• 关键词: Glycotechnology; Enzymatic Synthesis; Transglycosylation; Oligosaccharide Unit; Glycosidase; Glycoconjugate; Glycopolymer; Lectin; Glycosidase; Glycoconjugate; Glycopolymer; オリゴ糖鎖; ゴ糖鎖工学; 分子認識; 酵素合成

There is high currentinterest in developing synthetic routes to oligosaccharides involved in glycoconjugates. We established three synthetic routes of di- and trisaccharides, which are involved in glycoconjugate, through glycosidase-mediated transglycosylation. These synthetic oligosaccharides would be used for preparing glycopolymer as probes in carbohydrate-protein interaction studies.Firstly, Manbeta1-4GlcNAcbeta1-4GlcNAc, which is the common structural feature at the core region of most of the N-glycoproteins, was regioselectively synthesized from mannotriose and N,N'-diacetylchitobiose utilizing beta-D-mannanase from Aspergillus niger. Secondary, Galbeta1-3GalNAcalpha-OC_6H_4NO_2-p, which is a mimic unit at carbohydrate region of mucin type, was regioselectively synthesized from lactose and p-nitorpheny1 N-acetyl-a-galactosaminide uitilizing porcine testes beta-D-galactosidase-mediated transglycosylation^<1)>. Finally, Lacto-N-triose II glycoside, which is the common trisacccharide unit of glycosphingolipids, with two beta-(1-6)-linked isomers were synthesized through N-acetylglucosaminyl transfer by beta-N-acetylhexosaminidase from Nocardia orientalis. In this case, when an inclusion complex of p-nitrophenyl lactoside acceptor with beta-cyclodextrin was used, the regioselectivity of glycosidase-catalyzed formation of trisaccharide glycoside was substantially changed. It resulted not only in a significant increase of an overall yield of transfer products, but also in the proportion of the desired compound (GlcNAcbeta1-3Galbeta1-4Glcbeta-OC_6H_4NO_2-p). Furthermore, glycopolymers containing a series of beta-galactosyl-disaccharides were synthesized^<3)> and used as model in oligosaccharide-lectin interaction analysis. Their interaction with various lectins was investigated by double diffusion test and the inhibition of hemagglutinating activity of lectins. These water-soluble glycopolymers were shown to be an useful tool as probes of carbohy drate recognition.

目前，人们对开发糖缀合物中所涉及的寡糖的合成路线非常感兴趣。我们建立了三条通过糖苷酶介导的转糖基化反应合成糖缀合物中所涉及的二硫代和三硫代的路线。首先，利用黑曲霉（Aspergillusniger）的β-D-甘露聚糖酶，以甘露三糖和N，N '-二乙酰壳二糖为原料，区域选择性地合成了Manbeta 1 - 4GlcNAc β 1 -4GlcNAc，这是大多数N-糖蛋白核心区域的共同结构特征。其次，利用猪睾丸β-D-半乳糖苷酶介导的转糖基化反应，以乳糖和对硝基苯基N-乙酰-α-氨基半乳糖苷为原料，区域选择性地合成了粘蛋白糖区模拟单元Galbeta 1 -3GalNAcalpha-OC_6 H_4 NO_2-p.最后，利用Nocardia orientalis的β-N-乙酰氨基己糖苷酶，通过N-乙酰氨基葡萄糖转移，合成了具有两个β-（1-6）-连接异构体的鞘糖脂的常见三糖单元乳糖-N-丙糖II糖苷。在这种情况下，当使用对硝基苯基乳糖苷受体与β-环糊精的包合络合物时，糖苷酶催化形成三糖糖苷的区域选择性基本上改变。这不仅显著提高了转移产物的总收率，而且提高了目标产物（GlcNAc β 1 - 3Gal β 1 - 4Glc β-OC_6 H_4 NO_2-p）的比例。此外，合成了含有一系列β-半乳糖基-二糖的糖共聚物^<3）>，并将其用作寡糖-凝集素相互作用分析的模型。用双向扩散试验和凝集素的血凝抑制试验研究了它们与各种凝集素的相互作用。这些水溶性糖聚合物被证明是一个有用的工具，作为探针的碳水化合物识别。

项目成果

Y.Matashira et al.: "Enzymic synthesis of Lacto-N-triose II and its analogues" Glycoconjugate J.12. 664-671 (1995)

Y.Matashira 等人：“Lacto-N-triose II 及其类似物的酶合成”Glycoconjugate J.12。

K.Kobayashi et al.: "Synthesis of poly N-acetyl-β-lactosaminide-carring acrylamide via a chemical-enzymatic hybrid process" Methods in Enzymology. 242. 226-235 (1994)

K. Kobayashi 等人：“通过化学-酶混合工艺合成携带聚 N-乙酰基-β-乳糖胺的丙烯酰胺”，酶学方法 242. 226-235 (1994)。

Yoshiharu Matahira: "Enzymic Synthesis of Lacto-N-triose II and Its Positional Analogs" Glycoconjugate J.,accepted for publication.

Yoshiharu Matahira：“Lacto-N-triose II 及其位置类似物的酶合成”Glycoconjugate J.，接受出版。

K.Kobayashi et al.: "Chemoenzymatic synthesis of a glycopolymer carring clustered-N-acetyl-β-lactosaminide moieties" J. Carbohydr. Chem.13. 753-766 (1994)

K. Kobayashi 等人：“携带簇状 N-乙酰基-β-乳糖胺部分的糖聚合物的化学酶合成”J. CarboHydr. 753-766 (1994)。

Kazukiyo Kobayashi: "Chemo-Enzymatic Synthesis of a Glycopolymer Carring Clustered-N-acety1-β-Lactosaminide" J.Carbohydr.Chem.13(5). 753-766 (1994)

Kazukiyo Kobayashi：“携带簇状 N-乙酰基 1-β-乳糖酰胺的糖聚合物的化学酶法合成”J.CarboHydr.Chem.13(5) (1994)。