重组灵芝免疫调节蛋白的纯化及其性质

Purification and Properties of Recombinant Ganoderma Lucidum Immunoregulatory Protein

采用超滤浓缩、强阴离子交换、疏水作用和凝胶色谱等方法,对毕赤酵母表达的rGlip进行分离和纯化,对离子交换色谱中rGlip与固相结合的最佳pH值进行了考察,并对纯化产物的活性进行了鉴定.rGlip在215nm处有强的紫外吸收,经激光解析电离时间飞行质谱鉴定其相对分子量为12722,经反相液相色谱鉴定纯度≥97%.设计rGlip的疏水作用色谱,有效地去除色素.凝血实验结果表明,rGlip可以凝集绵羊血红细胞,但对人血A,B,AB和O型等红细胞无凝集作用,有类似凝集素的生物学活性.

Recombinant Ganoderma lucidum immunoregulatory protein （rGlip） was purified from fermentation. Purification was carried out by ultrafiltration, anion exchange chromatography（ AIEC ） , hydrophobic interaction chromatography（HIC）, size exclusion chromatography（SEC）. We used Hitrap Q sepharose column to measure the optimal pH value of anion exchange chromatography and detect the absorption value of protein at 215, 254, 280 nm. The intensity absorption of rGlip was found at 215 nm. Hemagglutination reactionno aggregation was observed between any types of human red blood cells, the positive activity was seen in the presence of sheep blood red cells. The optimal pH of AIEC enhanced the yield of initial purification, HIC was designed for lyophobic domains of rGlip, and end product（rGlip） of purification procedures possessed aggregation activity of sheep red blood cells.