High-density Escherichia coli cultivation process for hyperexpression of recombinant porcine growth hormone.
Fermentation studies were performed on an Escherichia coli culture that carries a recombinant plasmid composed of an ampicillin-resistant gene, a temperature-regulated pL promoter, and a porcine pituitary cDNA sequence coding for growth hormone. The objective was to achieve high cell density while maintaining the specific expression level of recombinant porcine growth hormone (r-pGH) observed in shake flasks. At a specific expression level of 20% of total cell protein, the cell density of a glucose-limited fed-batch process reached 38 units of OD600 in 14 h, compared to flask cultivation, which resulted in only 1.4 units of OD600 in the same period. The observed critical fermentation conditions for maximal expression included (1) limiting glucose concentration below 1 g l-1 throughout the fed-batch growth and induction phases, (2) keeping postinduction temperature at 42 degrees C for 5-7 h, and (3) maintaining a postinduction growth rate around 0.17-0.21 h-1.
Microbiology and Fermentation Division, Development Center for Biotechnology, Taipei, Taiwan, Republic of China.
H C
HC
Chen
C F
CF
Hwang
D G
DG
Mou
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D003470:Culture Media; D004926:Escherichia coli / Q000254:growth & development; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering / Q000379:methods*; D013006:Growth Hormone / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013552:Swine
D003470:Culture Media; D011994:Recombinant Proteins; D013006:Growth Hormone