High-density Escherichia coli cultivation process for hyperexpression of recombinant porcine growth hormone

Abstract

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 OD 600 in 14 h, compared to flask cultivation, which resulted in only 1.4 units of OD 600 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°C for 5–7 h, and (3) maintaining a postinduction growth rate around 0.17–0.21 h -1.