化工学报 ›› 2003, Vol. 54 ›› Issue (12): 1724-1728.

• 生物化学工程、制药、食品和天然产物加工 • 上一篇    下一篇

基于氧碳平衡的重组巴氏毕赤酵母碳源代谢分析

郭美锦; 储炬; 庄英萍; 杭海峰; 张嗣良   

  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),上海 200237
  • 出版日期:2003-12-25 发布日期:2003-12-25

CARBON SOURCE METABOLISM ANALYSIS FOR RECOMBINANT Pichia pastoris IN CHEMOSTAT BASED ON OXYGEN AND CARBON BALANCES

GUO Meijin;CHU Ju;ZHUANG Yingping;HANG Haifeng;ZHANG Siliang   

  • Online:2003-12-25 Published:2003-12-25

摘要: 基于氧元素和碳元素平衡对重组巴氏毕赤酵母(Pichia pastoris)的不同碳源代谢进行了分析.结果表明:以甘油为惟一碳源,当比生长速率由0.064h-1增大到0.257h-1时,用于细胞生长的甘油代谢比速率由1.36mmol•g-1•h-1增大到5.46mmol•g-1•h-1,而用于细胞维持代谢流比例降低了10.4%,同时甘油用于合成代谢和分解代谢的比速率均增大,但是分解代谢流比例逐渐下降,而合成代谢流比例却升高了5.9%;以甲醇为碳源进行表达重组人血清白蛋白恒化培养时,当比生长速率由0.019h-1增大至0.046h-1时,用于产物重组人血清白蛋白形成的比速率呈先增大后减少的趋势,在比生长速率为0.027h-1时最大,为0.31mmol•g-1•h-1,占甲醇总代谢流的13.2%.

关键词:

氧碳元素平衡, 重组巴氏毕赤酵母, 代谢流分析, 恒化培养, 重组人血清白蛋白

Abstract: Glycerol metabolism and methanol metabolism in genetically engineered Pichia pastoris were analyzed respectively in chemostat cultivation based on balances of oxygen and carbon elements.With the cell specific growth rate increasing from 0.064h-1 to 0.257h-1,the specific consumption rate of glycerol for cell growth metabolism increased from 1.36mmol•g-1•h-1 to 5.46mmol•g-1•h-1,whereas the percentage of glycerol metabolic flux for cell maintenance decreased from 14.4% to 4.0% when glycerol was the sole carbon source. The percentage of assimilation flux of glycerol increased by 5.9% while the percentage of catabolism flux of glycerol decreased by 5.9% , although the specific rates of assimilation and the specific rates of catabolism became larger.The recombinant human serum albumin was expressed in genetically engineered Pichia pastoris when methanol was used as substrate in continuous culture with specific growth rate increasing from 0.019h-1 to 0.046h-1.The largest specific consumption rate of methanol for target protein production was 0.31mmol•g-1•h-1,which was only 13.2% of all methanol metabolic flux at μ=0.027h-1.

Key words:

氧碳元素平衡, 重组巴氏毕赤酵母, 代谢流分析, 恒化培养, 重组人血清白蛋白