甘油相控制策略对重组毕赤酵母表达瑞替普酶(reteplase)的影响  

Effects of glycerol feeding strategies on production of recombinant reteplase from a methylotrophic yeast 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5期33-37,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：研究了甘油补料策略对毕赤酵母表达reteplase(rPA)的发酵过程中细胞的生长和rPA表达的影响。通过将对甘油补料速率由10g/L.h-1增大到20g/L.h-1,细胞比生长速率,甘油的比消耗速率,甘油得率等都得到极大提高。而且,诱导期细胞生长,甲醇消耗和rPA的生成速率都增加,rPA的最大表达量从140.2mg/L增加到199.5mg/L。此另外,甘油补料时间也是影响rPA表达的重要因素,甘油补料时间短,细胞密度小,表达rPA少,甘油补料时间长,细胞密度增大,rPA的表达速率也增加,但是到诱导后期,细胞死亡率增大,蛋白酶释放增加,rPA的降解增强。Effects of glycerol feeding strategies on the cell growth and recombinant reteplase（rPA） expression by the Pichia pastoris were studied. By increasing the glycerol feeding rate from 10 g/L· h^- 1 to 20 g/L· h^- 1 , not only the specific growth rate, specific glycerol comsuption rate and glycerol yield in glycerol phase were improved , but also the higher specific growth rate, specific methanol comsuption rate and rPA formation rate during induction phase were achieved. The maximum rPA was increased from 140.2mg/L to 199.5mg/L. Furthermore, the glycerol fed-batch phase period had a significant effect on rPA productivity. With the shorter glycerol fed-batch phase, the biomass became lower and the small quantity of rPA was expressed. While with the elongation of glycerol fed-batch phase, the biomass became higher, thus increasing the methanol comsuption and rPA expression. However, in the later production phase, the cell viability de- creased and the protease activity increased markedly, which led to a significant degradation of rPA.

关 键 词：毕赤酵母 RETEPLASE 甘油 

分 类 号：TQ920[轻工技术与工程—发酵工程]