Co-overexpression of RspAB Improves Recombinant Protein Production in Escherichia coli

Weikert, Christian; Canonaco, Fabrizio; Sauer, Uwe; Bailey, James E.

doi:10.1006/mben.2000.0163

Cited by 18 publications

(13 citation statements)

References 27 publications

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“…Similar cell physiology was observed in the ST8 strain, in which transcription of rspAB was activated, but that of rpoS was not affected. These results are not consistent with those of a previous report [58]. However, some other factors, such as elevated level of AI-2 signaling pathway in ST8 strain, could involve in the transcriptional upregulation of RspAB, because it was reported that AI-2 is related to osmotic stress and RpoS regulations [59,60].…”

Section: Stress-induced Responsecontrasting

confidence: 98%

“…It has been demonstrated that RspAB induces degradation of homoserine lactone (HSL), which affects the expression of RpoS [57]. E. coli, harboring rspAB expressing vector, featured reduced acetate production and increased recombinant protein yield [58]. Similar cell physiology was observed in the ST8 strain, in which transcription of rspAB was activated, but that of rpoS was not affected.…”

Section: Stress-induced Responsementioning

confidence: 92%

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Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Jung

Lim

et al. 2019

Microb Cell Fact

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Background: Most microorganisms have evolved to maximize growth rate, with rapid consumption of carbon sources from the surroundings. However, fast growing phenotypes usually feature secretion of organic compounds. For example, E. coli mainly produced acetate in fast growing condition such as glucose rich and aerobic condition, which is troublesome for metabolic engineering because acetate causes acidification of surroundings, growth inhibition and decline of production yield. The overflow metabolism can be alleviated by reducing glucose uptake rate. Results: As glucose transporters or their subunits were knocked out in E. coli, the growth and glucose uptake rates decreased and biomass yield was improved. Alteration of intracellular metabolism caused by the mutations was investigated with transcriptome analysis and 13 C metabolic flux analysis (13 C MFA). Various transcriptional and metabolic perturbations were identified in the sugar transporter mutants. Transcription of genes related to glycolysis, chemotaxis, and flagella synthesis was downregulated, and that of gluconeogenesis, Krebs cycle, alternative transporters, quorum sensing, and stress induced proteins was upregulated in the sugar transporter mutants. The specific production yields of value-added compounds (enhanced green fluorescent protein, γ-aminobutyrate, lycopene) were improved significantly in the sugar transporter mutants. Conclusions: The elimination of sugar transporter resulted in alteration of global gene expression and redirection of carbon flux distribution, which was purposed to increase energy yield and recycle carbon sources. When the pathways for several valuable compounds were introduced to mutant strains, specific yield of them were highly improved. These results showed that controlling the sugar uptake rate is a good strategy for ameliorating metabolite production.

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Section: Stress-induced Responsecontrasting

confidence: 98%

Section: Stress-induced Responsementioning

confidence: 92%

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Jung

Lim

et al. 2019

Microb Cell Fact

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“…Nevertheless, the idea that RspA may degrade HSL is consistent with the recent observation that RspA-overproducing strains indeed seem to have increased homoserine and decreased HSL levels (U. Sauer, personal communication). Unexpectedly, these strains actually show elevated S levels, which would not seem consistent with HSL being an inducer for rpoS (226).…”

Section: Small Molecules That Influence Rpos Transcriptionmentioning

confidence: 73%

Signal Transduction and Regulatory Mechanisms Involved in Control of the σ^S(RpoS) Subunit of RNA Polymerase

Hengge‐Aronis

2002

Microbiol Mol Biol Rev

851

791

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SUMMARY The σS (RpoS) subunit of RNA polymerase is the master regulator of the general stress response in Escherichia coli and related bacteria. While rapidly growing cells contain very little σS, exposure to many different stress conditions results in rapid and strong σS induction. Consequently, transcription of numerous σS-dependent genes is activated, many of which encode gene products with stress-protective functions. Multiple signal integration in the control of the cellular σS level is achieved by rpoS transcriptional and translational control as well as by regulated σS proteolysis, with various stress conditions differentially affecting these levels of σS control. Thus, a reduced growth rate results in increased rpoS transcription whereas high osmolarity, low temperature, acidic pH, and some late-log-phase signals stimulate the translation of already present rpoS mRNA. In addition, carbon starvation, high osmolarity, acidic pH, and high temperature result in stabilization of σS, which, under nonstress conditions, is degraded with a half-life of one to several minutes. Important cis-regulatory determinants as well as trans-acting regulatory factors involved at all levels of σS regulation have been identified. rpoS translation is controlled by several proteins (Hfq and HU) and small regulatory RNAs that probably affect the secondary structure of rpoS mRNA. For σS proteolysis, the response regulator RssB is essential. RssB is a specific direct σS recognition factor, whose affinity for σS is modulated by phosphorylation of its receiver domain. RssB delivers σS to the ClpXP protease, where σS is unfolded and completely degraded. This review summarizes our current knowledge about the molecular functions and interactions of these components and tries to establish a framework for further research on the mode of multiple signal input into this complex regulatory system.

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“…Successful evolutionary engineering examples exist with a variety of industrially important microorganisms, such as Escherichia coli, where physiological properties such as stress resistance (Weikert et al, 1997), recombinant protein production (Weikert et al, 2000), metabolic activity, and lactic acid production (Fong et al, 2005) were improved, Acetobacter aceti with improved acetate resistance (Steiner & Sauer, 2003), Bacillus subtilis with improved plasmid stability (Fleming et al, 1988), and Actinobacillus succinogenes (Roca et al, 2010) and Mannheimia succiniciproducens with enhanced succinate production. However, many other evolutionary engineering studies are with the baker's yeast S. cerevisiae to improve a variety of physiological and production properties of this microorganism.…”

Section: Inverse Metabolic Engineering and Its Applicationsmentioning

confidence: 99%

Evolutionary engineering of Saccharomyces cerevisiae for improved industrially important properties

et al. 2011

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This article reviews evolutionary engineering of Saccharomyces cerevisiae. Following a brief introduction to the 'rational' metabolic engineering approach and its limitations such as extensive genetic and metabolic information requirement on the organism of interest, complexity of cellular physiological responses, and difficulties of cloning in industrial strains, evolutionary engineering is discussed as an alternative, inverse metabolic engineering strategy. Major evolutionary engineering applications with S. cerevisiae are then discussed in two general categories: (1) evolutionary engineering of substrate utilization and product formation and (2) evolutionary engineering of stress resistance. Recent developments in functional genomics methods allow rapid identification of the molecular basis of the desired phenotypes obtained by evolutionary engineering. To conclude, when used alone or in combination with rational metabolic engineering and/or computational methods to study and analyze processes of adaptive evolution, evolutionary engineering is a powerful strategy for improvement in industrially important, complex properties of S. cerevisiae.

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Co-overexpression of RspAB Improves Recombinant Protein Production in Escherichia coli

Cited by 18 publications

References 27 publications

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Signal Transduction and Regulatory Mechanisms Involved in Control of the σ^S(RpoS) Subunit of RNA Polymerase

Evolutionary engineering of Saccharomyces cerevisiae for improved industrially important properties

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Co-overexpression of RspAB Improves Recombinant Protein Production in Escherichia coli

Cited by 18 publications

References 27 publications

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Signal Transduction and Regulatory Mechanisms Involved in Control of the σS(RpoS) Subunit of RNA Polymerase

Evolutionary engineering of Saccharomyces cerevisiae for improved industrially important properties

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Product

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Signal Transduction and Regulatory Mechanisms Involved in Control of the σ^S(RpoS) Subunit of RNA Polymerase