温度对巴斯德毕赤酵母表达瑞替普酶(reteplase)的影响  

Effects of temperature on expression of reteplase in Pichia pastoris

作　　者：方曙光[1] 储炬[1] 黄立[1] 庄英萍[1] 张嗣良[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室国家生化工程技术研究中心,上海200237

出　　处：《工业微生物》2007年第4期11-15,共5页Industrial Microbiology

基　　金：国家"十五"高新技术发展计划(863计划)项目(2002AA217021);国家重大科技专项项目(2002AA2Z3451)

摘　　要：研究了温度对巴斯德毕赤酵母表达瑞替普酶(reteplase,rPA)的影响。结果发现,在BMMY摇瓶培养条件下,诱导温度在20℃、25℃,rPA表达量较高,分别是30℃诱导条件下(18.2mg/L)的1.4倍和1.34倍。在高密度发酵过程中,降低诱导温度(20℃、25℃),rPA的表达量在诱导表达84h时达到最高,分别为207.9mg/L和199.5mg/L,比30℃诱导条件下分别提高了35%和30%。通过对细胞活性、蛋白酶和AOX酶活性分析发现,降低温度不仅提高了发酵过程中酵母活细胞率,降低了蛋白酶的活性,减少了rPA的降解,而且提高了AOX酶活,增强了rPA的表达。The effects of temperature on the production of recombinant reteplase（rPA） in Pichia pastoris were investigated. The results demonstrated that in BMMY shake-flask culture, the expression of rPA at lower temperatures （20℃, 25℃; ） was 1.4 and 1.34 times higher than that at 30℃;, respectively. This was verified in high cell density cultures. The yield of rPA at lower temperatures （20℃; ,25℃; ） were 207.9mg/L and 199.5mg/L, respectively, both much higher than that cultured at high temperature （30℃, 153.2 mg/L）. The analysis of protease, cell viability and AOX activity showed that cultures at lower temperature not only led to a reduced cell death ratio and hence a lower amount of host cell proteases in the supematant, but also significant increase of AOX activity. Those might be benefit for rPA production.

关 键 词：毕赤酵母 RETEPLASE 降解 温度 

分 类 号：Q786[生物学—分子生物学]