Sigma factors as potential targets to enhance recombinant protein expression

Abstract

The transcriptional factors control the expression of many genes and represent an important layer of complexity in cell factories. However, the effect of individual sigma factor deletions from a biomanufacturing perspective has not been addressed. In this contribution, growth and oxygen consumption of E. coli BW25113 strains with individual inactivation of each sigma factor were characterized under various conditions. The deletion of rpoD has been reported to be lethal for E. coli. However, the evaluated rpoD mutant did not exhibit major growth defects in the mineral medium and is explained because the rpoD gene is duplicated in this strain. Recombinant protein was expressed at three different induction levels in a mineral and LB media. The fliA mutant was the best producer in the mineral medium, whereas the rpoD mutant overperformed the other strains in LB medium. This suggest that a lower rpoD gene dosage is positive for the performance of the cell factory in complex medium. In cultures at 20C, the rpoS mutant exhibited the greatest recombinant expression. To our knowledge, this is the first systematic study evaluating the potential of sigma factor deletion for improving recombinant protein production.