School of Pharmaceutical Sciences, South-Central University for Nationalities, Wuhan 430074, China.
State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, China.
Mar Drugs. 2020 May 29;18(6):286. doi: 10.3390/md18060286.
Mild acid hydrolysis is a common method for the structure analysis of fucosylated glycosaminoglycan (FG). In this work, the effects of acid hydrolysis on the structure of FG from S. variegatus (SvFG) and the reaction characteristic were systemically studied. The degree of defucosylation (DF) and molecular weights (Mw) of partial fucosylated glycosaminoglycans (pFs) were monitored by H NMR and size-exclusion chromatography, respectively. The kinetic plots of DF, degree of desulfation (DS) from fucose branches, and degree of hydrolysis (DH) of the backbone are exponentially increased with time, indicating that acid hydrolysis of SvFG followed a first-order kinetics. The kinetic rate constants k, k, and k were determined to be 0.0223 h, 0.0041 h, and 0.0005 h, respectively. The structure of the released sulfated fucose branches (FucS) from SvFG and HfFG (FG from ) was characterized by 1D/2D NMR spectroscopy, suggesting the presence of six types of fucose: α/β Fuc2S4S, Fuc3S4S, Fuc3S, Fuc4S, Fuc2S, and Fuc. The Fuc3S4S was more susceptible to acid than Fuc2S4S, and that the sulfate ester in position of O-2 and O-3 than in O-4 of fucose. The structure characteristic of pF18 indicated the cleavage of backbone glycosidic bonds. The APTT prolonged activity reduced with the decrease of the DF and Mw of the pFs, and became insignificant when its DF was 87% with Mw of 3.5 kDa.
温和酸水解是分析岩藻糖基化糖胺聚糖（FG）结构的常用方法。本工作系统研究了酸水解对花尾胡椒鲷 FG（SvFG）结构的影响及反应特性。采用 1H NMR 和凝胶渗透色谱分别监测部分岩藻糖基化糖胺聚糖（pFs）的去岩藻糖基化程度（DF）和分子量（Mw）。DF、糖链中来自岩藻糖支链的去硫酸化程度（DS）和主链水解程度（DH）的动力学曲线随时间呈指数增加，表明 SvFG 的酸水解遵循一级动力学。确定动力学速率常数 k、k 和 k 分别为 0.0223 h、0.0041 h 和 0.0005 h。通过 1D/2D NMR 光谱对 SvFG 和 HfFG（FG 来自）释放的硫酸化岩藻糖支链（FucS）的结构进行了表征，表明存在六种类型的岩藻糖：α/β Fuc2S4S、Fuc3S4S、Fuc3S、Fuc4S、Fuc2S 和 Fuc。Fuc3S4S 比 Fuc2S4S 更容易受到酸的影响，而且在岩藻糖的 O-2 和 O-3 位的硫酸酯比 O-4 位更容易受到酸的影响。pF18 的结构特征表明其主链糖苷键被切断。pFs 的 DF 和 Mw 降低，APTT 延长活性降低，当 DF 为 87%、Mw 为 3.5 kDa 时，APTT 延长活性变得不显著。
