Fed‐batch operation of recombinant Escherichia coli containing trp promoter with controlled specific growth rate

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Abstract
A feb-batch operation for the production of bovine somatotropin (bST) under the control of tryptophan promoter in Escherichia coli was investigated. The plasmid used contains a two-cistron system and altered codon usage for higher expression of bST. Specific growth rate is an important parameter in the fermentation, because it affects the production of growth-inhibitory organic acids and the expression of recombinant protein. The feeding rate was adjusted to keep the specific growth rate constant in this study. The variable growth yield expressed as a function of time was used for the calculation of the feeding rate. The growth yield decreases during the fermentation as product expression is induced. The specific growth rate was well controlled; however, intracellular bST concentration decreased at high cell concentrations. This is considered to be due to degradation by proteases. The decrease was prevented by an exponential feeding of the yeast extract as an organic nitrogen source. © 1994 John Wiley & Sons, Inc.