Development of Glyco-medicines Using Mutant Enzyme of Microbial Endoglycosidase and Establishment of Its Preparation Method

利用微生物糖苷内切酶突变酶开发糖类药物及其制备方法的建立

• 批准号: 20380052
• 负责人: YAMAMOTO Kenji
• 金额: $ 12.23万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20380052/
• 关键词: エンドグリコシダーゼ; 糖転移活性; オキサゾリン化合物; 部位特異的変異酵素; 化学-酵素合成; 糖鎖医薬品; インフルエンザウィルス; 感染阻害剤; substrate assisted catalysis; サブスタンスP; PAMP12; リボヌクレアーゼB; シアロ糖鎖; インフルエンザウイルス; 変異酵素

The endo-β-N-acetylglucosaminidase from Mucor hiemalis isolated from soil sample is unique in that it can transfer en bloc the oligosaccharide of glycoproteins onto different acceptors, and thereby it enzymatically generates diverse glycoconjugates. We performed mutational studies to obtain the mutant of the enzyme which had significantly diminished hydrolysis activity but had a high transglycosylation activity. We have found that some mutant could efficiently exhibit transglycosylation activity using synthetic sugar oxazoline. Therefore, we used the mutant enzyme to synthesize an efficient binding inhibitor for influenza virus, which is composed of sialyloligosaccharides on chitosan. This compound showed sufficient inhibitory activity against influenza virus infection.

从土壤样品中分离的冬夜毛霉（Mucor hiemalis）的内切-β-N-乙酰氨基葡萄糖苷酶（endo-β-N-acetylglucosaminidase）是一种独特的酶，它能将糖蛋白中的寡糖整体转移到不同的受体上，从而酶促生成不同的糖缀合物。我们进行了突变研究，以获得酶的突变体，其水解活性显著降低，但具有高的转糖基化活性。我们发现一些突变体可以有效地利用合成糖恶唑啉表现出转糖基活性。因此，我们利用该突变酶合成了一种有效的流感病毒结合抑制剂，它是由唾液酸寡糖在壳聚糖上组成的。该化合物对流感病毒感染显示出足够的抑制活性。

项目成果

Glycosynthases Enable a Highly Efficient Chemoenzymatic Synthesis of N-Glycoproteins Carrying Intact Nature N-Glycans

糖合酶能够高效化学酶合成携带完整天然 N-聚糖的 N-糖蛋白

• 发表时间: 2009
• 期刊: Journal of The American Chemical Society 131
• 作者: W.Huang;et al.
• 通讯作者: et al.

Simple Synthesis of Influenza Virus Binding-Inhibitor Composed of Only Sugar Chains from Natural Compounds

从天然化合物中简单合成仅由糖链组成的流感病毒结合抑制剂

• 发表时间: 2008
• 作者: 梅村舞子;ら
• 通讯作者: ら

Mutants of Mucor hiemlis Endo-β-N-acetylglucosaminidase Show Enhanced Transglycosylation and Glycosynthase-like Activities

冬毛霉内切-β-N-乙酰氨基葡萄糖苷酶突变体表现出增强的转糖基化和糖合酶样活性

• 发表时间: 2008
• 期刊: The Journal of Biological Chemistry 283
• 作者: M. Umekawa;et al.
• 通讯作者: et al.

Design of a sialylglycopolymer with a chitosan backbone having efficient inhibitory activity against influenza virus infection.

• DOI: 10.1021/jm8000967
• 发表时间: 2008-07
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Myco Umemura;M. Itoh;Y. Makimura;Kohji Yamazaki;M. Umekawa;Ayano Masui;Y. Matahira;M. Shibata;H. Ashida;Kenji Yamamoto

Efficient Transfer of Sialo-oligosaccharides onto Proteins by Combined Use of a Glycosynthase-like Mutant of Mucor hiemalis Endoglycosidase and Synthetic Sialo-complex-type Sugar Oxazoline.

通过结合使用冬毛霉内切糖苷酶的糖合酶样突变体和合成的唾液酸复合物型糖恶唑啉，将唾液酸低聚糖有效转移到蛋白质上。

• 发表时间: 2010
• 期刊: Biochim. Biophys. Acta 1800
• 作者: M.Umekawa;T.Higashiyama;Y.Koga;T.Tanaka;M.Noguchi;A.Kobayashi;S.Shoda;W.Huang;L.-X.Wang;H.Ashida;K.Yamamoto
• 通讯作者: K.Yamamoto