嗜麦芽寡养单胞菌(DR-929)纤溶酶的发酵条件优化及其分离纯化  

Optimization on Fermentation and Purification of the Fibrinolytic Enzyme from Stenotrophomonas maltophilia

作　　者：王高学[1] 梁朝军[1] 黄海洪[1] 王建福[1] 原居林[1] 

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100

出　　处：《中国生物工程杂志》2007年第9期47-52,共6页China Biotechnology

基　　金：陕西省科技攻关资助项目(2006K02-G14-03)

摘　　要：研究了嗜麦芽寡养单胞菌（DR-929）纤溶酶的液体发酵条件及其分离纯化。最佳发酵条件为：可溶性淀粉2．0％，黄豆粉1．0％，酵母膏0．5％，NaCI1．0％，CaCl20．02％，MgS040．05％，种龄36h，发酵时间4d，初始pH8．0或9．0，温度25℃，装样量30ml，接种量5％或6％。采用发酵液离心除菌，25％-70％饱和度的硫酸铵沉淀，Phenyl FF（highsub）疏水层析，Q—SepharoseFF离子交换层析，Superdex 75凝胶过滤层析对活性成分分离纯化。用SDS—PAGE电泳对纯化效果进行检验，结果表明在SDS—PAGE中得到单一条带，分子量28．3kDa。最终纯化倍数和酶活回收率分别为271．5和24．5％。The optimization on liquid fermentation and purification of the fibrinolytic enzyme from Stenotrophomonas maltophilia（DR-929） was investigated. The results showed the best fermentation are amidulin 2.0%, soya flour 1.0%, yeast extract 0.5%, NaCI 1.0%, CaCI2 0.02%, MgSO4 0.05% , inoculum of 36 hours, fermental time 4d, initial pH 8.0 or 9.0, temperature 25 ℃, volume of media 30ml, volume of inoculum 5% or 6%. The purification process includes the following steps： removing cells by the centrifugation, 25% - 70% saturation ammonium sulfate precipitation, HIC with Phenyl FF（ high sub）, IEC with Q-Sepharose FF, gel filtration chromatography with Superdex 75. SDS-PAGE electrophoresis was used to examine the purification effect, and the results indicated that homogeneous strap in SDS-PAGE and has a molecular weight around 28.3kDa. The purification factor and activity recovery of the fibrinolytic enzyme are 271. 5 and 24. 5%, respectively.

关 键 词：嗜麦芽寡养单胞菌 纤溶酶 发酵条件 分离纯化 

分 类 号：Q814[生物学—生物工程]