Ishii Nozomi, Sano Kanae, Matsuo Ichiro
Graduate School of Science and Technology, Gunma University, 1-5-1, Tenjin-cho, Kiryu, Gunma 376-8515, Japan.
Graduate School of Science and Technology, Gunma University, 1-5-1, Tenjin-cho, Kiryu, Gunma 376-8515, Japan.
Bioorg Med Chem Lett. 2019 Jul 1;29(13):1643-1646. doi: 10.1016/j.bmcl.2019.04.039. Epub 2019 Apr 26.
We synthesized a fluorogenic probe with a high-mannose type heptasaccharide structure to detect the hydrolytic activity of endo-β-N-acetylglucosaminidase from Streptomyces plicatus (Endo-H). The heptasaccharide derivative (1) was labeled with an N-methylanthraniloyl group as a reporter dye at the branching point of the β-mannoside residue and 2,4-dinitrophenyl group as a quencher molecule at the reducing end, which was hydrolyzed by Endo-H, resulting in increased fluorescence intensity. Thus, Endo-H activities could be evaluated easily and quantitatively by measuring the fluorescence signal. Using both this probe (1) and a previously synthesized pentasaccharide probe, the hydrolysis activity of Endo-H and Endo-M were investigated. The results clearly showed a correlation with the substrate specificity of each enzyme.
我们合成了一种具有高甘露糖型七糖结构的荧光探针，用于检测褶皱链霉菌内切-β-N-乙酰氨基葡萄糖苷酶（Endo-H）的水解活性。七糖衍生物（1）在β-甘露糖苷残基的分支点处用N-甲基邻氨基苯甲酰基作为报告染料进行标记，并在还原端用2,4-二硝基苯基作为猝灭分子，该分子可被Endo-H水解，导致荧光强度增加。因此，通过测量荧光信号可以轻松、定量地评估Endo-H的活性。使用该探针（1）和先前合成的五糖探针，研究了Endo-H和Endo-M的水解活性。结果清楚地表明了与每种酶的底物特异性的相关性。
