Department of Biology & Guangdong Provincial Key Laboratory of Marine Biotechnology, College of Science, Shantou University, Shantou, Guangdong 515063, PR China.
Department of Mechanical Engineering, College of Engineering, Shantou University, Shantou, Guangdong 515063, PR China.
Carbohydr Polym. 2017 Oct 1;173:465-472. doi: 10.1016/j.carbpol.2017.06.023. Epub 2017 Jun 12.
Three separation methods for water-soluble polysaccharides from Spirulina platensis (PSP) were compared, including sevage deproteinization and column chromatography, ethanol-ammonium sulfate aqueous two-phase system (ATPS) and column chromatography, and one-step high speed counter-current chromatography (HSCCC) with ethanol-ammonium sulfate ATPS. ATPS was confirmed as an efficient alternative method of protein removal for PSP purification. For the HSCCC with ethanol-ammonium sulfate ATPS, the stationary retention reached to 50.8% at the optimized rotation speed and flow rate. Moreover, the yield of PSP purified by one-step HSCCC rose nearly five times to 12.45mg/g(dry algae powder), it was higher than PSP yield by two-step column chromatography separation methods. PSP purified by HSCCC has the same purity as PSP obtained by traditional methods, which was proved by a single symmetrical peak of purified PSP with molecular weight of 12.33kDa through gel chromatography. Purified PSP was an α-acidic polysaccharide, composed of major glucose, slight rhamnose and mannose, which were detected within GC and FT-IR spectra. The antioxidation activity experiment showed that HSCCC-purified PSP had strong scavenging effects on hydroxyl free radical and DPPH free radical.
三种分离方法从螺旋藻（PSP）的水溶性多糖进行了比较，包括sevage 脱蛋白和柱层析，乙醇-硫酸铵双水相系统（ATPS）和柱层析，以及一步高速逆流色谱（HSCCC）与乙醇-硫酸铵 ATPS。ATPS 被确认为蛋白质去除的有效替代方法 PSP 净化。HSCCC 与乙醇-硫酸铵 ATPS 时，固定相保留率达到 50.8%，在优化的转速和流速。此外，由一步 HSCCC 纯化 PSP 的产量增加了近五倍，至 12.45 毫克/克（干藻粉），它比两步柱层析分离方法的 PSP 产量高。HSCCC 纯化 PSP 具有相同的纯度，通过凝胶色谱纯化的 PSP 分子量为 12.33kDa，证明具有单一对称峰。纯化的 PSP 是一种α-酸性多糖，由主要葡萄糖、少量鼠李糖和甘露糖组成，在 GC 和 FT-IR 光谱中检测到。抗氧化活性实验表明，HSCCC 纯化 PSP 对羟基自由基和 DPPH 自由基有很强的清除作用。
