Role of the general stress response during strong overexpression of a heterologous gene in Escherichia coli.
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Role of the general stress response during strong overexpression of a heterologous gene in Escherichia coli. / Schweder, T; Lin, Hongying; Jürgen, B; Breitenstein, A; Riemschneider, S; Khalameyzer, V; Gupta, A; Büttner, K; Neubauer, P.
In: APPL MICROBIOL BIOT, Vol. 58, No. 3, 3, 2002, p. 330-337.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Role of the general stress response during strong overexpression of a heterologous gene in Escherichia coli.
AU - Schweder, T
AU - Lin, Hongying
AU - Jürgen, B
AU - Breitenstein, A
AU - Riemschneider, S
AU - Khalameyzer, V
AU - Gupta, A
AU - Büttner, K
AU - Neubauer, P
PY - 2002
Y1 - 2002
N2 - 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.
AB - 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.
M3 - SCORING: Zeitschriftenaufsatz
VL - 58
SP - 330
EP - 337
JO - APPL MICROBIOL BIOT
JF - APPL MICROBIOL BIOT
SN - 0175-7598
IS - 3
M1 - 3
ER -
