豆豉溶栓酶工程菌的发酵条件及重组溶栓酶的分离纯化  

Fermentation of Douchi Fibrinolytic Enzyme Gene Engineering Strain and the Purification of Recombinant Enzyme

作　　者：肖璐[1] 张仁怀[1] 张义正[1] 

机构地区：[1]四川大学生命科学学院四川省分子生物学及生物技术重点实验室,成都610064

出　　处：《食品与发酵工业》2005年第1期66-69,73,共5页Food and Fermentation Industries

摘　　要：对豆豉溶栓酶工程菌株WB DFE5进行了摇瓶发酵条件的研究 ,确定了发酵的优化条件为 :可溶性淀粉 2 % ,酪蛋白 2 % ,大豆蛋白胨 0 5 % ,酵母粉 0 15 % ,K2 HPO40 2 5 % ,KH2 PO40 0 5 % ,CaCl2 0 0 2 % ,MgSO40 0 5 % ,pH7 0。通过硫酸铵分段盐析、离子交换和凝胶过滤 ,从 1L工程菌株WB DFE5的发酵液中分离纯化出 12 5mg重组豆豉溶栓酶 ,酶的比活为 5 918 7IU/mg。In order to increase production of recombinant Douchi fibrinolytic enzyme(rDFE),the optimal fermentation conditions of recombinant Bacillus subtilise WB-DFE5 were determined. The compositions of culture medium are 2% soluble starch, 2% casein, 0.5% soybean peptone, 0.15% yeast extract, 0.25% K 2HPO 4, 0.05% KH 2PO 4, 0.02% CaCl 2, 0.05% MgSO 4, pH7.0. rDFE was purified by methods including ammonium sulfate fractional precipitation, SP-Sepharose FF, DEAE-Sepharose FF ion-exchang and Sephadex G-75 chromatography.12.5 mg of purified product was obtained from one liter of culture medium and the enzyme spectific activity is 5918.7IU/mg.

关 键 词：豆豉 溶栓酶 大豆蛋白 可溶性淀粉 分离纯化 酪蛋白 发酵条件 酶工程 过硫酸铵 摇瓶发酵 

分 类 号：TQ925[轻工技术与工程—发酵工程]