Yoshioka K, Takahashi H, Homma T, Saito M, Oh K B, Nemoto Y, Matsuoka H
Department of Biotechnology, Tokyo University of Agriculture and Technology, Japan.
Biochim Biophys Acta. 1996 Feb 9;1289(1):5-9. doi: 10.1016/0304-4165(95)00153-0.
A novel fluorescent derivative of glucose was synthesized by reacting D-glucosamine and NBD-Cl. The TLC analysis of the reaction mixture showed the generation of a single spot with intense fluorescence (lambda Ex = 475 nm, lambda Em = 550 nm). The obtained novel fluorescent product, which was identified as 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) by 1H-NMR and FAB-MS spectrometries, was applied to the assessment of the glucose uptake activity of Escherichia coli B. 2-NBDG accumulated in living cells and not in dead cells. The uptake of 2-NBDG was competitively inhibited by D-glucose and not by L-glucose, which suggested the involvement of the glucose transporting system in the uptake of 2-NBDG. 2-NBDG taken into the cytoplasma of E. coli cells was supposedly converted into another derivative in the glucose metabolic pathway.
通过使D - 葡糖胺与NBD - Cl反应合成了一种新型的葡萄糖荧光衍生物。反应混合物的TLC分析显示产生了一个具有强荧光的单一斑点（激发波长λEx = 475 nm，发射波长λEm = 550 nm）。通过1H - NMR和FAB - MS光谱鉴定，所得到的新型荧光产物为2 -（N -（7 - 硝基苯并 - 2 - 恶唑 - 1,3 - 二唑 - 4 - 基）氨基）-2 - 脱氧葡萄糖（2 - NBDG），并将其应用于评估大肠杆菌B的葡萄糖摄取活性。2 - NBDG在活细胞中积累，而不在死细胞中积累。2 - NBDG的摄取受到D - 葡萄糖的竞争性抑制，而不受L - 葡萄糖的抑制，这表明葡萄糖转运系统参与了2 - NBDG的摄取。进入大肠杆菌细胞质的2 - NBDG可能在葡萄糖代谢途径中转化为另一种衍生物。
