重组巴氏毕赤酵母恒化培养动力学及代谢迁移特性研究  

STUDIES ON CHARACTERISTICS OF KINETICS AND METABOLIC SHIFT OF GENETICALLY ENGINEERED YEAST PICHIA PASTORIS IN HIGH-DENSITY CHEMOSTAT CULTIVATION

作　　者：郭美锦[1] 吴康华[1] 庄英萍[1] 储炬[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

出　　处：《微生物学报》2001年第5期617-624,共8页Acta Microbiologica Sinica

基　　金：国家自然科学基金资助项目 ( 2 99760 1 3);教育部高等学校骨干教师资助计划资助项目

摘　　要：通过对甲醇营养型毕赤酵母基因工程菌以碳源甘油为限制性基质进行恒化培养动力学试验 ,结果认为 :(1 )细胞光密度与其干、湿重呈线性关系 ,当细胞光密度 (OD60 0 )为 1 0 0时细胞湿重 (WCW)为 1 2 8 3g L ,细胞干重 (WDW)则为 2 2 9g L ;(2 )基因工程菌P .pastoris的生长与限制性基质甘油残留浓度的关系符合Monod关系式 ,通过 1 μ对 1 S进行线性回归得 μmax=0 .366h- 1,Ks=0 .1 82 3g L ,经参数推导甘油最大菌体得率系数YG =0 .54g g ,菌体维持生长消耗底物系数m =0 .0 0 69g (g·h) ;氧最大菌体系数YX O2 =30 .96g moL ,菌体维持生长时消耗氧系数mO2 =0 .0 0 0 8mol (g·h) ,最适理论稀释速率Dm =0 .341h- 1;(3)从氨水的消耗速率和呼吸商 (RQ)的变化认为随着比生长速率 (μ)的增大 ,甘油代谢流从糖原异生和磷酸戊糖途径线性地向糖酵解和三羧酸循环途径进行代谢迁移 。Kinetics and metabolic shift of DNA recombinant yeast Pichia pastoris was studied in high\|density chemostat cultivation in which the glycerol was used as a limited substrate.The experimental results showed that (1) cell optical density ( OD 600 ) of Pichia pastoris was linear with its dry cell weight (DCW) and wet cell weight (WCW),and reached 100 OD 600 =128.3g WCW /L or100 OD 600 =22.9g DCW /L;(2)The relationship of specific growth rate ( μ ) and residual concentration of glycerol is accorded with Monod Equation,to obtain μ=μ max S/(Ks+S),where μ max =0.366h -1 ,Ks=0.1823g/L.The maximum cell yield of glycerol substrate Y\-G=0.54g/g,growth maintaining coefficient m=0.0069g/(g·h),The cell yield of oxygen Y X/O2 =30.96g/moL,and the optimum theoretical dilution rate Dm=0.341h -1 by parameter inference;(3)With specific growth rate ( μ ) increasing,glycerol metablic flux shifts linearly from glycogenesis and hexosephosphate pathways to glycolysis and tricarboxylic acid cycle pathways on basis of variations of consumption rate of NH\-3·H\-2O and respiratory quotient value.

关 键 词：甲醇营养型毕赤酵母 恒化培养 代谢迁移 动力学 重组人血清白蛋白 

分 类 号：Q939.9[生物学—微生物学]