木瓜蛋白酶法与超滤膜法提取柿子多糖最佳条件的筛选  

Screening of optimum conditions for extraction of total polysaccharides from persimmon fruits by papain deproteinization and ultra- filtration membranes methods

作　　者：谢玲[1] 郑林希 夏志兰[1] 钟晓红[1] 

机构地区：[1]湖南农业大学园艺园林学院,长沙410128

出　　处：《果树学报》2017年第1期106-114,共9页Journal of Fruit Science

基　　金：湖南农业大学园艺学开放课题(2013YYX017)

摘　　要：【目的】降低传统化学试剂提取而带来的毒性大风险,试用简便、温和、环保又高效的木瓜蛋白酶法、超滤膜法等分离纯化柿子总多糖提取物(TPs PF)。【方法】以当季成熟‘牛心柿’果实为试材,探讨了木瓜蛋白酶脱蛋白过程中不同处理工艺对TPs PF保留率和蛋白质去除率的影响,并进一步选择MWCO为200/10 ku的超滤膜截留收集分子质量10~200 ku的目标多糖组分,以超滤膜通量和TPs PF截留率为评价指标筛选出超滤工艺条件。【结果】TPs PF木瓜蛋白酶法脱蛋白优化工艺参数:酶用量1.0%,温度60℃,处理时间1.5 h,p H值6或7。其中对TPs PF保留率影响较大的因素是处理时间,而对蛋白质去除率影响较大的则是p H值。超滤膜富集分子质量10~200 ku的TPs PF的优化工艺条件:压力0.3 MPa,温度30℃,初始多糖浓度20 g·L^(-1),最后得到TPs PF截留率为(45.82±2.37)%。【结论】试验中的脱蛋白酶法、超滤法等生物分离方法操作简便,温和并获得了较高的提取率,适合于TPs PF的分离提取,并为下一步药理作用的评价提供了物质基础。[Objective] TPsPF （total polysaccharides from persimmon fruits） is known to have a variety of pharmacological activities. Extraction of the TPsPF by traditional chemical reagents would potentially have some toxic risk. The extraction of the TPsPF by biological and physical methods would be valuable for the reason of the safety of the extracts. Therefore, papain deproteinization and ultra-filtration membranes were employed for extracting the TPsPF in present study. [Methods ] The flesh of persimmon was defated by chloroform and methanol （3 ： 1） at 60 ℃, and the crude TPsPF was extracted under reflux with alcohol. Then, different conditions of papain deproteinization by using removal rate of protein and retention rate of TPsPF as indexes. The effect of the enzyme content, temperature, processing time and pH value were investigated using the method of single factor test. After the treatments of papain deproteinization and decolorization of active carbon, three kinds of membranes with different molecular weight cut-off （MWCO） were used to filter the TPsPF. The TPsPF of 10-200 ku were enriched by using two membranes with MWCO （200 ku, 10 ku）, meanwhile the effect of operating pressure, the polysaccharide concentration and temperature on ultra-filtration membrane flux and the retention rate of TPsPF were studied and the optimal conditions were selected. [ Results ] According to the results of retention rate of TPsPF and removal rate of protein, the conditions of papain deproteinization process were optimized as follows： enzyme content of 1.0%, at 60 22, pH 6 （or 7） and 1.5 h. When enzyme content was 1.0%, the highest retention rate of TPsPF was 92% and removal rate of protein was about 50%. The removal rate of protein reached the peak when enzyme content was 1.5%. Regarding the cost, we preferred to use 1.0% of enzyme. When the temperature varied from 30 22 to 60 ℃,there were no apparent change of the retention rate of the TPsPF and the removal rate of protein. However, the retention rate of

关 键 词：柿子总多糖 脱蛋白酶法 木瓜蛋白酶 超滤膜 截留分子质量 

分 类 号：S665.2[农业科学—果树学]