Production of granulocyte-macrophage colony stimulating factor (GM-SCF)by high cell density fermentation of secretory recombination Escherichia Coli

High cell density fermentation production of recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF)was studied in an Escherichia coli secretory expression system. By using glycerol instead of glucose as a carbon source and feeding in a specific fed-batch mode and by controlling the temperature at about 30deg C and maintaining the dissolved oxygen level at 20 approx 25% saturation, cell density increased from 30 to 91.8 g~(-1),the production of recombinant GM-CSF remained high and the fermenation time was reduced from 45 to 30 h. Recombination GM-CSF was purified by hydrophobic column-molecular sieve-ion exchange column chromatography. The expression of recombiant GM-CSF was not affected by fermentation methods,sieve-ion exchange column chromatography. The expression of recombinant GM-CSF was not affected by fermentation methods, the production of GM-CSF reached 28.04% (about 1.68 g 1~(-1)) and 27.51%(about 1.64 g 1~(-1)) of total somatic proteins for high cell density and normal fermenatation, respectively. The corresponding value for specific activity of GM-CSF was 2.6*10~7, and 2.4*10~7,respectively.

作 者：

Zhang,,Xue-Wu

刊 名：

Process Biochemistry 

年，卷(期)：

1999Vol. 34 No.1 Jan 1999("") 

分类号：

 

关键词：

GM-CSF  E.cell  High cell density fermentation  Purification

正文语种：

eng 

基金项目：