鳆发光杆菌荧光素酶的提取、分离与纯化  

Purification of luciferase from P.Ieiognathi

作　　者：俎聪[1] 李明[1] 鲁晓东[1] 王静雪[1] 

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：《食品工业科技》2013年第12期243-245,250,共4页Science and Technology of Food Industry

基　　金：国家自然科学基金(31071540);山东省自然科学基金(ZR2011CQ024)

摘　　要：建立了从鳆发光杆菌中提取纯化荧光素酶的方法。鳆发光杆菌25℃培养12h,经过超声破碎、硫酸铵沉淀等步骤后得到粗酶液,再通过DEAE Sepharose CL-6B、DEAE Sephadex A-50进一步纯化,得到电泳纯的荧光素酶,通过SDS-PAGE测得其有两个亚基,分子量分别为36、41ku。经过硫酸铵沉淀以及两步层析后,荧光素酶的纯化倍数达到100倍以上,证明该实验过程具有较好的纯化效果,为荧光素酶性质的研究和商业化应用提供了技术支持。A method to purify luciferase from P.leiognathi was proposed. P.leiognathi was incubated for 12h at 25℃. Afterwards,luciferase was purificated by sonication,solid ammonium sulfate,DEAE Sepharose CL-6B and DEAE Sephadex A-50 columns. Electrophoresis indicated luciferase had two subunits and their molecular weights were 36ku and 41ku respectively. By salting out and two steps chromatography,more than 100-fold purified luciferase was obtained. Results showed that the method was feasible for purification of luciferase and provided technical support for its properties study and commercialization.

关 键 词：鳆发光杆菌 荧光素酶 纯化 

分 类 号：TS201.1[轻工技术与工程—食品科学]