文献类型：期刊文献

中文题名：基因工程菌Pichia pastoris高密度培养条件研究

英文题名：STUDIES ON CONDITIONS OF CELL HIGH DENSITY CULTIVATION FOR GENETICALLY ENGINEERED METHYLOTROPHIC PICHIA PASTORIS

作者：郭美锦[1];吴康华[1];杭海峰[1];储炬[1];庄英萍[1];张嗣良[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室

年份：2001

卷号：28

期号：3

起止页码：6

中文期刊名：微生物学通报

外文期刊名：Microbiology China

收录：CSTPCD;;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

基金：国家自然科学基金!资助项目 (No. 2 99760 1 3 );教育部高等学校骨干教师资助项目&&

语种：中文

中文关键词：基因工程菌;毕赤酵母;高密度培养

外文关键词：Methylotrophic Pichia pastoris,High density cultivation,Recombinant human serum albumin

摘要：基因工程菌pichiapastoris最佳种子培养基为添加 4mL/LPTMl的BMGY培养基 ;全合成高密度摇瓶培养基是甘油 4% ,(NH4) 2 SO41 0g/L ,CaSO40 93g/L ,K2 SO41 8 2g/L ,MgSO4·7H2 O 1 4 9g/L ,0 1mol/L磷酸缓冲液 (pH =6 0 )配制 ,培养 2 6h后细胞密度OD60 0 可达到 65。经SDS PAGE电泳图谱分析 ,甲醇诱导培养 72h结果 :1 2h有重组人血清白蛋白表达 ,2 4h达到最大。此全合成摇瓶培养基与批补料发酵培养基相类似 ,有利于指导发酵罐上发酵培养。
Recently the methylotrophic Pichia pastoris,with many advantages such as high expression,stable genetics and protein secretion,has been used extensively as a host expressing heterologous proteins.The optimal seed medium among seven media MM,MD,MGY,BMGY,YMPD,YMPGy and MGyB is BMGY with addition of 4mL/L PTM1 The high density synthesized medium in shake flask culture is as follows: Glycerol 4%(NH 4) 2SO 4 10g/L,CaSO 4 0.93g/L,K 2SO 4 18 2g/L,MgSO 4·7H 2O 14 9g/L,add 0.1mol/L PBS(pH=6 0) to 1 0 liter.The harvested yeast cell optical density (OD 600 ) reached 65 or more after 26 hours cultivation.By analysis of SDS\|PAGE,the results of Pichia pastoris culture by methanol inducement for 72 hours showed that the expression of recombinant human serum albumin had been achieved by methanol inducement after 12 hours,and the mount of targeted protein reached the summit after 24 hours inducement by methanol.The high density synthesized medium in shake flask culture in this experiment,which is similar to the mediums of batch fermentation and batch\|fed fermentation,is benefit to direct the P.pastoris fermentation in the fermentor.