摘要
利用大孔树脂吸附结合硅胶柱层析技术对发酵液中多杀菌素的分离纯化进行了研究。首先考察了提取溶剂、发酵液pH和料液比对浸提多杀菌素的影响；然后对7种不同极性的大孔吸附树脂进行了静态和动态吸附性能的研究，考察了不同吸附、解吸条件对大孔吸附树脂性能的影响，最后对硅胶柱层析法精制多杀菌素的工艺条件进行了探索。结果表明：当发酵液pH为8．O，料液比1：3（g：v）时，乙醇可高效浸提多杀菌素；大孔吸附树脂DMll对多杀菌素的静态吸附量和解吸率分别为12508μg/g（湿树脂）和93．47％；大孔树脂动态吸附最佳pH为9．0，最佳载样比为多杀菌素质量（mg）：树脂体积（mL）=2．5：1；采用70％-95％乙醇梯度洗脱，多杀菌素洗脱率为97．5％；多杀菌素精制时硅胶柱以石油彬乙酸乙酯／甲醇（2：1：0．2和1：1：0．25）为洗脱剂进行分段洗脱。该工艺得到的多杀菌素回收率为67．25％，纯度为90．58％。
Extraction and purification of spinosad from fermentation broth has been studied by macroporous resin adsorption supported with silica gel chromatography technology. First, the influences of the type of extraction sol-vents, extraction solvents volume and pH in spinosad extraction process were investigated. Second, the static and dy-namic adsorption expressions of seven varieties of macroporous resins for spinosad extraction were studied and the op-timum macroporous resin was selected in present process. Finally, the silica gel chromatography technology was ex- plored for spinosad purification. The results showed that ethanol had ability to extract the spinosad efficiently on the condition of the pH of fermentation broth 8.0, the ratio of material and solvent was 1：3 （g/v）. DMll was selected as the optimum macroporous resin to extract and separate spinosad on the experimental condition; the adsorption and desorption ratio of DM11 macroporous resin for spinosad was 12 508 μg/g（ wet resin） and 93.47% respectively. The. maximum adsorption capacity of DM11 macroporous resin for spinosad could be reached, at pH 9.0. The optimal ratio of spinosad and macroporous resin for adsorption was 2.5：1 （rag： mL）. When 70%-95% ethanol was utilized as eluant, the desorption capacity reached 97.5 %. The silica gel column chromatography was adopted for further purifi- cation of spinosad by gradient elution using a mixture of petroleum ether： ethyl acetate ： methanol （2 ： 1 ： 0.2 and 1：1：0.25 ）, and the highest purity of 90.58% was obtained under these conditions.
出处
《中国粮油学报》
EI
CAS
CSCD
北大核心
2014年第3期95-101,共7页
Journal of the Chinese Cereals and Oils Association
基金
"十二五"国家科技支撑计划(2011BAD03B02-2)
2013年"农业科技成果转化资金项目"(2013GB 24490638)
关键词
多杀菌素
大孔吸附树脂
硅胶层析
分离
纯化
spinosad, macroporous resin, silica gel column, separation, purification
