蛹虫草类胡萝卜素酶法提取和TLC法纯化研究  

Study on Enzymatic Extraction and TLC Purification of Cordyceps militaris Carotenoids

作　　者：卜义明 江晓路[1] 孟菲 吕秀霞 BU Yi-ming;JIANG Xiao-lu;MENG Fei;LYU Xiu-xia(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003

出　　处：《中国食用菌》2020年第5期30-36,42,共8页Edible Fungi of China

基　　金：青岛市“十三五”海洋经济创新发展示范城市项目(2016QD003)。

摘　　要：为了开发一种条件温和、安全环保的方法来分离纯化水溶性蛹虫草类胡萝卜素,通过试验筛选得到辅助提取蛹虫草类胡萝卜素的最佳酶为脂肪酶,并优化形成最佳工艺。蛹虫草子实体粉与水1∶4 (m∶v)混匀,加入0.1%脂肪酶,于pH 8~8.5,50°C条件下酶解30 min;将酶解液的乙醇浓度调至60%,室温下静置提取90 min;离心收集上清液。用该工艺提取1次蛹虫草类胡萝卜素的含量为(1 747.8±17.6)μg·g-1,显著高于直接浸提法的[(652.3±24.9)μg·g-1]和超声辅助提取法[(661.5±41.9)μg·g-1],P<0.01。通过试验筛选得到蛹虫草类胡萝卜素TLC纯化法的最佳展开剂配比为:0.5%乙酸水溶液∶乙醇∶正丁醇∶乙酸乙酯=12∶8∶24∶36。用该展开剂能分离形成4个清晰的条带。经HPLC对比分析,4个条带的溶出液在出峰时间和顺序上与文献报道的4种蛹虫草类胡萝卜素Cordyxanthin-Ⅰ、Cordyxanthin-Ⅱ、Cordyxanthin-Ⅲ、Cordyxanthin-Ⅳ分别一一对应,峰面积百分比分别为76.10%、95.90%、97.96%、81.53%。In order to develop a mild,safe and environmentally friendly method for the separation and purification of water-soluble Cordyceps militaris carotenoids,the lipase was selected as the best enzyme to assist in extracting C.militaris carotenoids,and the best process was as follow:the C.militaris fruiting body powder and water were mixed at a ratio of 1:4(m:v),then 0.1% lipase was added.The enzymatic hydrolysis reaction was carried out for 30 min in the condition of pH 8-8.5 and 50℃,the ethanol concentration of the enzymatic hydrolysate was adjusted to 60%,then it was placed at room temperature for 90 min,the supernatant was collected by centrifugation.The content of C.militaris carotenoids was(1747.8±17.6)μg·g^-1after extracting once by this process,which was significantly higher(P<0.01)than the direct extraction method of(652.3±24.9)μg·g^-1. and the ultrasound-assisted extraction method of(661.5±41.9)μg·g^-1.The optimum TLC developing solvent ratio of C.militaris carotenoids was 0.5%aqueous acetic acid:ethanol:n-butanol:ethyl acetate is 12:8:24:36.Four kinds of strips were clearly separated by the developing solvent.By HPLC analysis,the four strips of the eluate were correspond to the four C.militaris carotenoids(Cordyxanthin-I,Cordyxanthin-II,Cordyxanthin-III,Cordyxanthin-IV)reported in the literature,and the peak area percentages are 76.10%,95.90%,97.96%,81.53%,respectively.

关 键 词：蛹虫草类胡萝卜素 酶法提取 TLC纯化 

分 类 号：S646.1[农业科学—蔬菜学]