小球藻热水提取物功能成分的活性跟踪分离

Bioassay-guided isolation of functional components from hot water extract of Chlorella pyrenoidosa

摘要:

本研究旨在以自由基清除作用和促巨噬细胞增殖作用为导向，对小球藻热水提取物 (CPE) 的分离纯化产物进行活性跟踪，以确定CPE的主要功能成分。采用高压热水提取、Sevag除蛋白、乙醇沉淀和超滤的方法得到CPE粗多糖，然后通过DEAE52离子交换层析柱和Sephadex G-100对CPE粗多糖组分进一步分离纯化。研究结果表明，通过上述方法对CPE进行分离纯化，得到3种高活性成分PS-1-4-2、PS-1-3-2和PS-2-3-3，经凝胶渗透色谱GPC分析得知其分子质量分别为3.97×104 Da、2.28×104 Da和4.1×103 Da。活性跟踪结果表明，CPE能够清除自由基并促进巨噬细胞Ana-1细胞生长，主要是由于其多种活性组分共同发挥作用，清除自由基作用的功能成分主要集中于PS-1-3、PS-1-4、PS-2-3和PS-2-4，促Ana-1细胞增殖作用的功能成分主要集中于PS-1-3、PS-1-4和PS-2-3。本研究确立了CPE主要功能成分的活性筛选手段，并获得3种新型功能成分，可用于指导小球藻高附加值产品的开发，从而进一步推动微藻能源的产业化进程，实现“高附加值微藻产品、微藻能源与微藻固碳”一体化的示范作用。

Abstract:

The main functional ingredients of hot water extract of Chlorella pyrenoidosa (CPE) were investigated through a bioassay-guided fractionation based on free radical scavenging and macrophage proliferation effects. The main functional ingredients of CPE were polysaccharides (PS) that were isolated by high pressure extraction, Sevag method, ethanol precipitation and ultrafiltration separation. Crude polysaccharides were further separated and purified by ion exchange chromatography DEAE52 and size exclusion chromatography Sephadex G-100. The purified fractions were analyzed by gel permeation chromatography. Molecular weights of the purified fractions PS-1-4-2, PS-1-3-2 and PS-2-3-3 were 3.97×104, 2.28×104 and 4.1×103 Dalton, respectively. Bioassay-guided fractionation results indicated that CPE could remove free radicals and promote Ana-1 cells proliferation, mainly due to its various components working together. The components of free radicals scavenging mainly concentrated in PS-1-3, PS-1-4, PS-2-3 and PS-2-4. The components of Ana-1 proliferation mainly concentrated in PS-1-3, PS-1-4 and PS-2-3. This study established the activity screening method of main functional component from CPE, and got three new functional ingredients. It can be used to guide the development of high value products, further promote the industrialization process of microalgae energy, and realize microalgae ‘high value products, microalgae energy and microalgae carbon’ integration of exemplary role.