澳洲坚果多糖脱蛋白方法研究  

Deproteinization of macadamia nut polysaccharide

作　　者：许良[1] 叶丽君[1] 黄雪松[1] 

机构地区：[1]暨南大学理工学院食品科学与工程系,广州510632

出　　处：《食品科技》2014年第12期206-211,共6页Food Science and Technology

基　　金：公益性行业农业科研专项(201303077)

摘　　要：为得到纯化的澳洲坚果多糖,采用热水提取多糖,并对其进行分步醇沉。以蛋白脱除率和多糖损失率为指标,比较Sevage法、TCA法、碱性蛋白酶法和碱性蛋白酶-Sevage联用法对60%-90%醇沉粗多糖脱除蛋白的效果,再利用DEAE-Sepharose Fast Flow及Superdex 200柱层析进一步纯化多糖。结果表明,碱性蛋白酶-Sevage联用法为多糖脱蛋白最佳方法,其最优条件为碱性蛋白酶酶用量1.0%（v/v）、酶解温度50℃、酶解时间3 h,再经Sevage法处理一次,蛋白脱除率达92.3%,多糖损失率为39.9%。随后经DEAE-Sepharose Fast Flow及Superdex 200柱层析纯化得到澳洲坚果多糖纯品（MIPS2a）,紫外和红外扫描表明MIPS2a不含蛋白且具备一般多糖类物质的光谱特征,达到了多糖纯化的目的。The macadamia nut kernel polysaccharide was extracted by hot water, and the supernatant wasprecipitated by stepwise alcohol. Protein removal rate and polysaccharide loss rate were used to evaluate the effectiveness of Sevage＇s method, TCA method, alkaline protease method and alkaline proteaseSevage＇s method in deproteinizing 60%-90% precipitated crude polysaccharide. Then the preliminary deproteinized polysaccharide was loaded onto DEAE-Sepharose Fast Flow and Superdex 200 column respectively for further deproteinization. The results showed that the alkaline protease-Sevage＇s method was the best way for removing protein in polysaccharide, and the the optimum condition was using 1%（v/v） alkaline protease, hydrolyzing 3 h at the temperature of 50 ℃, followed by Sevage＇s method. A protein removal rate of 92.3% and a polysaccharide loss rate of 39.9% were achieved under such conditions. The preliminary deproteinized polysaccharide could be completely purified by collecting the main eluting peak of DEAE-Sepharose Fast Flow and Superdex 200 clomn. The pure polysaccharide, named MIPS2 a, had no protein characteristic absorption showed in UV. In addition, MIPS2 a had infrared spectral characteristic of common polysaccharides, further indicating that the pure polysaccharide was obtained.

关 键 词：澳洲坚果 多糖 脱蛋白 

分 类 号：TS255.6[轻工技术与工程—农产品加工及贮藏工程]