基于氧碳平衡的重组巴氏毕赤酵母碳源代谢分析  

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

作　　者：郭美锦[1] 储炬[1] 庄英萍[1] 杭海峰[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),上海200237

出　　处：《化工学报》2003年第12期1724-1728,共5页CIESC Journal

基　　金：国家自然科学基金资助项目 (No 2 99760 13 );教育部高等学校骨干教师资助项目~~

摘　　要：基于氧元素和碳元素平衡对重组巴氏毕赤酵母 (Pichiapastoris)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 0 6 4h-1增大到 0 2 5 7h-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 4 6mmol·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 0 19h-1增大至 0 0 4 6h-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 0 2 7h-1时最大 ,为 0 31mmol·g-1·h-1,占甲醇总代谢流的 13 2 % .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.

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 

分 类 号：TQ933[化学工程]