Effect of growth rate on stability and gene expression of recombinant plasmids during continuous and high cell density cultivation of Escherichia coli TG1.
Continuous and fed-batch cultures of recombinant Escherichia coli TG1 were carried out in order to study plasmid stability and recombinant product formation at different specific growth rates. The aprotinin::beta-galactosidase gene (Ap::lacZ) was placed under the control of two different promoter/repressor systems, the PLac/lacI (pPLac8) and the lambda PL/cIts857 (pPL6) system. The chemically (0.5 mM IPTG) induced gene expression exhibited higher product activity and plasmid stability than the thermally (40 degrees C) induced expression. In fed-batch cultivations with the more stable E. coli TG1(pPLac8) a special feeding strategy allowed bacterial growth with a constant growth rate mu for several hours up to high cell densities. The cloned gene product activity was noticeably effected by the specific growth rate and the cell density at the moment of induction. In particular, the enzyme activities passed a pronounced maximum value in dependence of the set growth rate. The results indicate that fed-batch cultivation strategies are well suited to produce recombinant gene products.
Hellmuth
K
K
GBF-Gesellschaft für Biotechnologische Forschung, Biochemical Engineering, Braunschweig, Germany.
Korz
D J
DJ
Sanders
E A
EA
Deckwer
W D
WD
D004926:Escherichia coli / Q000254:growth & development* / Q000378:metabolism; D015870:Gene Expression; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis*; D001616:beta-Galactosidase / Q000096:biosynthesis
D011994:Recombinant Proteins; D001616:beta-Galactosidase