链霉菌H03发酵液中多糖的分离纯化及其抗氧化活性的研究

Study on Isolation,Purification,Identification and Anti-oxidative Activity of Polysaccharide from Broth of Streptomyces H03

本实验对链霉菌H03发酵产物进行了分离纯化与鉴定,并利用邻苯三酚自氧化法检测此多糖消除超氧阴离子自由基的效果、用比色法测定多糖对羟自由基诱导红细胞溶血、肝微粒体脂质过氧化反应的影响等来研究其抗氧化活性。结果表明,利用Savag法去掉蛋白质、透析法去掉小分子物质以及醇沉后得到的粗提物,经Sephadex G-100凝胶柱层析可得到纯品,采用紫外光谱、红外光谱以及化学方法证明这种物质是一种多糖;而且这种链霉菌多糖具有较强的抗氧化活性,能显著消除超氧阴离子自由基、抑制羟自由基对细胞膜的破坏而引起的溶血和抑制肝微粒体脂质过氧化产物的形成。

A water soluble substance was isolated from the broth of Streptomyces H03 by method of ethanol precipitation, removing the protein by Savag method and gel permeation column chromatography. The ultraviolet, infrared spectrum and chemical analysis results showed that it is a novel polysaccharide. And the effects of anti-oxidative activities of the polysaccharide were studied in vitro. Scavenging superoxide anion activity was assayed by self-oxidation method of pyrogallol, and effects of the polysaccharide on red blood cell hemolysis and lipid peroxidation induced by hydroxyl free radical were analysed by colorimetric assay. Results showed that superoxide anion can be scavenged by the polysaccharide significantly, which exhibited a significant potency in inhibiting murine erythrocyte hemoyisis（p 〈 0.01）and lipid peroxidation in mouse liver homogenate（p 〈 0.01）.