CH9 产生的胞外多糖的纯化、初步结构与抗肿瘤活性
Purification, Preliminary Structure and Antitumor Activity of Exopolysaccharide Produced by CH9.
机构信息
Key Laboratory of Food Nutrition and Safety, Ministry of Education, Department of Food Engineering and Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China.
出版信息
Molecules. 2018 Nov 6;23(11):2898. doi: 10.3390/molecules23112898.
In the present study, the preliminary structure and in vitro antitumor activity of three exopolysaccharides (EPSs) from CH9 were investigated. Then, three purified fractions of EPS-1a, EPS-2a, and EPS-3a were obtained by chromatography using DEAE-52 cellulose and Sephadex G-100, respectively. The average molecular weight of EPS-1a, EPS-2a, and EPS-3a, were 1.80 × 10⁶, 1.06 × 10⁶ and 1.05 × 10⁶. The monosaccharide composition of EPS-3a was dramatically different from the others. The EPS-1a and EPS-2a were mainly composed of mannose, in a ratio of 69.82% and 57.09%, respectively, while EPS-3a was mainly composed of glucose (63.93%), without mannose. In addition, the surface morphology observed suggested that there were protein particles on the sugar chain of EPS-3a and EPS-3a was a protein-containing polysaccharide. Furthermore, EPS-3a exhibited higher antitumor activity against human liver cancer HepG2 cells in vitro. The antitumor activity of EPS-3a in HepG2 cells was associated with cell apoptosis. HE staining and Hoechst 33342 staining showed that with the treatment of EPS-3a, HepG2 cells had typical morphological changes. Flow cytometry analysis showed that the cell cycle was arrested at G0/G1 phase.
在本研究中，研究了 CH9 中的三种胞外多糖（EPS）的初步结构和体外抗肿瘤活性。然后，通过 DEAE-52 纤维素和 Sephadex G-100 层析分别得到三种纯化的 EPS-1a、EPS-2a 和 EPS-3a 级分。EPS-1a、EPS-2a 和 EPS-3a 的平均分子量分别为 1.80×106、1.06×106 和 1.05×106。EPS-3a 的单糖组成与其他两种明显不同。EPS-1a 和 EPS-2a 主要由甘露糖组成，比例分别为 69.82%和 57.09%，而 EPS-3a 主要由葡萄糖（63.93%）组成，不含甘露糖。此外，观察到的表面形态表明 EPS-3a 的糖链上存在蛋白质颗粒，并且 EPS-3a 是一种含有蛋白质的多糖。此外，EPS-3a 在体外对人肝癌 HepG2 细胞表现出更高的抗肿瘤活性。EPS-3a 在 HepG2 细胞中的抗肿瘤活性与细胞凋亡有关。HE 染色和 Hoechst 33342 染色显示，随着 EPS-3a 的处理，HepG2 细胞具有典型的形态变化。流式细胞术分析显示细胞周期被阻滞在 G0/G1 期。
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