Role of the general stress response during strong overexpression of a heterologous gene in Escherichia coli.
The strong overexpression of heterologous genes in Escherichia coli often leads to inhibition of cell growth, ribosome destruction, loss of culturability, and induction of stress responses, such as a heat shock-like response. Here we demonstrate that the general stress response, which is connected to the stress response regulator sigmas (sigma38, rpoS gene product), is suppressed during strong overproduction of a heterologous alpha-glucosidase. The mRNA levels of the rpoS and osmY stress genes drastically decrease after induction of the strong overexpression system. It is shown that an rpoS mutation causes a significant loss of cell viability after induction of the expression system. Furthermore, it is demonstrated that an E. coli c/pP mutant, which could be suggested to improve heterologous protein production, is not a good production host if a tac-promoter is used to control the expression of the recombinant gene. Data from this study suggest that the overexpression of the alpha-glucosidase was greatly decreased by sigma factor competition in the clpP mutant, due to the increased sigmas level in this mutant background.
Schweder
T
T
Institute of Microbiology, Ernst-Moritz-Arndt-University, Greifswald, Germany.
Lin
H Y
HY
Jürgen
B
B
Breitenstein
A
A
Riemschneider
S
S
Khalameyzer
V
V
Gupta
A
A
Büttner
K
K
Neubauer
P
P
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism; D000520:alpha-Glucosidases / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins; D012808:Sigma Factor; C069567:sigma factor KatF protein, Bacteria; D000520:alpha-Glucosidases; C063842:dnaK protein, E coli