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.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
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)
关键词
嗜麦芽寡养单胞菌
纤溶酶
发酵条件
分离纯化
Stenotrophomonas maltophilia Fibrinolytic enzyme Fermentation condition Purification
