Zhejiang University, College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Fuli Institute of Food Science, Zhejiang R & D Center for Food Technology and Equipment, Hangzhou 310058, China; Center for Biotechnology & Interdisciplinary Studies and Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Biotechnology Center 4005, Troy, NY 12180, United States.
Zhejiang University, College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Fuli Institute of Food Science, Zhejiang R & D Center for Food Technology and Equipment, Hangzhou 310058, China.
Carbohydr Polym. 2019 Oct 15;222:115025. doi: 10.1016/j.carbpol.2019.115025. Epub 2019 Jun 27.
Fucosylated chondroitin sulfate (FCS) oligosaccharides of specific molecular weight have shown potent anticoagulant activities with selectivity towards intrinsic factor Xase complex. However, the preparation of FCS oligosaccharides by traditional methods requires multiple purification steps consuming large amounts of time and significant resources. The current study focuses on developing a method for the rapid preparation of FCS oligomers from sea cucumber Pearsonothuria graeffei having 6-18 saccharide residues. The key steps controlling molecular weight (Mw) and purity of these FCS oligomers were evaluated. Structural analysis showed the resulting FCS oligomers were primarily l-Fuc3,4diS-α1,3-d-GlcA-β1,3-(d-GalNAc4,6diS-β1,4-[l-Fuc3,4diS-α1,3-]d-GlcA-β1,3-)d-anTal-ol4,6diS (n = 1˜5) accompanied by partial de-fucosylation and/or de-sulfation. In vitro and in vivo experiments demonstrate that these FCS oligomers selectively inhibit intrinsic factor Xase complex and exhibit remarkable antithrombotic activity without hemorrhagic and hypotension side effects. This method is suitable for large-scale preparation of FCS oligosaccharides as clinical anticoagulants.
岩藻依聚糖硫酸寡糖（FCS）具有特定分子量，对内在因子 Xase 复合物具有很强的抗凝活性，且具有选择性。然而，传统方法制备 FCS 寡糖需要经过多次纯化步骤，耗费大量时间和大量资源。本研究致力于从海参 Pearsonothuria graeffei 中快速制备具有 6-18 个糖基的 FCS 低聚物。评估了控制这些 FCS 低聚物分子量（Mw）和纯度的关键步骤。结构分析表明，得到的 FCS 低聚物主要为 l-Fuc3,4diS-α1,3-d-GlcA-β1,3-(d-GalNAc4,6diS-β1,4-[l-Fuc3,4diS-α1,3-]d-GlcA-β1,3-)d-AnTal-ol4,6diS（n=1˜5），同时伴有部分脱岩藻糖基化和/或脱硫酸基化。体内外实验表明，这些 FCS 低聚物选择性地抑制内在因子 Xase 复合物，具有显著的抗血栓形成活性，且无出血和低血压副作用。该方法适用于临床抗凝剂的 FCS 寡糖的大规模制备。
