径向流色谱脱除灵芝多糖中蛋白质和色素的工艺优化

摘要

径向流色谱是一种新型色谱分离纯化技术,在生物工程领域具有很大的应用潜力.文中利用径向流色谱法对灵芝粗多糖进行纯化,以脱除其中的蛋白质和色素.通过单因素及响应面试验,确定了纯化过程的最优化条件:上样流速2 mL/min,洗脱流速40 mL/min,上样量100 mL.在此条件下,蛋白脱除率、色素脱除率和多糖回收率分别达到80.35%、88.52%、85.06%.径向流色谱法与Sevag法、三氯乙酸法、活性炭法及双氧水法等传统化学方法比较的结果表明:径向流色谱脱蛋白、脱色及多糖保留效果均优于其他方法,且在处理过程中未使用有机试剂,突出了径向流色谱法高效、经济、安全、环保等优势.%Radial flow chromatography (RFC) was a new technique of chromatography purification, and has a great potential in biotechnology. The deproteinization and decoloration of polysaccharide from Ganoderma lucidium by radial flow chromatography were studied. The optimal conditions were determined by single factor experiments and response surface experiments: sample concentration 10 mg/mL, sample volume 100 mL, sample flow- rate 2 mL/min and elution flow - rate 40 mL/min. The deproteinization rate, decoloration rate and polysaccharide recovery rate under the optional chromatography condition were 80.35％ , 88.52％ and 85.06％, respectively. The purification efficiency of RFC was compared with sevag method, trichloroacetic acid method, activated carbon method and hydrogen perioxide method. The comparative results indicated that the purification by RFC were superior to other methods.None organic reagent in RFC treatment shows its advantages of high - efficiency, more economic, safe, and environmental friendly.