Dynamics of Escherichia coli at elevated temperatures: effect of temperature history and medium

Derlinden, Eva Van; Bernaerts, Kristel; Impe, Jan Van

doi:10.1111/j.1365-2672.2007.03592.x

Cited by 18 publications

(46 citation statements)

References 56 publications

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“…MG1655 cannot grow on LB plates at temperatures at or exceeding 46°C. This is consistent with reports that MG1655 grows very poorly in liquid LB at 46°C (58). Strain EVG1031 was sampled on day 77 when consistent and robust growth was seen in the Evolugator culture chamber at 46.9°C.…”

Section: Resultssupporting

confidence: 76%

“…However, while this study elegantly showed that serial batch culture could be used to improve the thermal properties of individual enzymes, no attempt was made to alter the overall thermal properties of the original organism (although alteration in individual enzymes could conceivably have a significant impact on global thermal properties). In a more salient example, Rudolph et al recently reported the use of experimental evolution over the course of 2 years of traditional serial batch culture to adapt Escherichia coli K-12 MG1655 to steadily increasing temperatures (49), resulting in a thermotolerant strain that could grow at 48.5°C, well above the T max of the parent strain (58). However, while they reported proteomic characterization of the resulting strain, they did not perform any sequencing to characterize the adaptive genotype.…”

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confidence: 99%

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Experimental Evolution of a Facultative Thermophile from a Mesophilic Ancestor

Blaby¹,

Lyons²,

Wroclawska-Hughes³

et al. 2012

Appl Environ Microbiol

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Experimental evolution via continuous culture is a powerful approach to the alteration of complex phenotypes, such as optimal/ maximal growth temperatures. The benefit of this approach is that phenotypic selection is tied to growth rate, allowing the production of optimized strains. Herein, we demonstrate the use of a recently described long-term culture apparatus called the Evolugator for the generation of a thermophilic descendant from a mesophilic ancestor (Escherichia coli MG1655). In addition, we used whole-genome sequencing of sequentially isolated strains throughout the thermal adaptation process to characterize the evolutionary history of the resultant genotype, identifying 31 genetic alterations that may contribute to thermotolerance, although some of these mutations may be adaptive for off-target environmental parameters, such as rich medium. We undertook preliminary phenotypic analysis of mutations identified in the glpF and fabA genes. Deletion of glpF in a mesophilic wild-type background conferred significantly improved growth rates in the 43-to-48°C temperature range and altered optimal growth temperature from 37°C to 43°C. In addition, transforming our evolved thermotolerant strain (EVG1064) with a wild-type allele of glpF reduced fitness at high temperatures. On the other hand, the mutation in fabA predictably increased the degree of saturation in membrane lipids, which is a known adaptation to elevated temperature. However, transforming EVG1064 with a wildtype fabA allele had only modest effects on fitness at intermediate temperatures. The Evolugator is fully automated and demonstrates the potential to accelerate the selection for complex traits by experimental evolution and significantly decrease development time for new industrial strains.

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Section: Resultssupporting

confidence: 76%

mentioning

confidence: 99%

Experimental Evolution of a Facultative Thermophile from a Mesophilic Ancestor

Blaby¹,

Lyons²,

Wroclawska-Hughes³

et al. 2012

Appl Environ Microbiol

View full text Add to dashboard Cite

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“…Previous studies showed that the addition of chemical chaperones (TMAO) could promote E. coli growth at elevated temperature, even up to 46°C (Gur et al 2002;Derlinden et al 2008). Ferrer et al (2003) reported that in vivo expression of the chaperonin Cpn60 and co-chaperonin Cpn10 isolated from a psychrophilic bacterium Oleipira antartica enabled the growth of the recombinant E. coli at temperature as low as 4°C, which indicated the possibility that the recruiting heterologous chaperones could extend the temperature ranges of growth of the cells.…”

Section: Discussionmentioning

confidence: 98%

Over-expression and characterization of recombinant prefoldin from hyperthermophilic archaeum Pyrococcus furiosus in E. coli

Chen

Yang

Zhang³

et al. 2009

Biotechnol Lett

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Prefoldin is a hexameric chaperone that has been identified in eukaryotic cells and in Archaea. E. coli cells over-expressing the prefoldin gene from the hyperthermophilic, archaeum, Pyrococcus furiosus, grew well at 48 degrees C while control cells were unable to grow above 46 degrees C. The isolated and purified Pfu-prefoldin (specially the beta subunit and the prefoldin) thermally protected the activity of lysozyme.

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“…Escherichia coli, being the most extensively studied bacterium and also a popularly utilized host cell for producing pharmaceutically important recombinant proteins, is known to be unable to grow at a temperature higher than 46.5°C (4)(5)(6). It has been widely reported that heterologous overexpression of certain exogenous molecular chaperones (7)(8)(9)(10)(11) or an endogenous transcriptional regulator (12) is able to significantly increase the viability of E. coli cells undergoing heat shock treatment at lethal temperatures (around 50°C).…”

mentioning

confidence: 99%

A Small Heat Shock Protein Enables Escherichia coli To Grow at a Lethal Temperature of 50 C Conceivably by Maintaining Cell Envelope Integrity

Ezemaduka

Shi

et al. 2014

Journal of Bacteriology

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It is essential for organisms to adapt to fluctuating growth temperatures. Escherichia coli, a model bacterium commonly used in research and industry, has been reported to grow at a temperature lower than 46.5°C. Here we report that the heterologous expression of the 17-kDa small heat shock protein from the nematode Caenorhabditis elegans, CeHSP17, enables E. coli cells to grow at 50°C, which is their highest growth temperature ever reported. Strikingly, CeHSP17 also rescues the thermal lethality of an E. coli mutant deficient in degP, which encodes a protein quality control factor localized in the periplasmic space. Mechanistically, we show that CeHSP17 is partially localized in the periplasmic space and associated with the inner membrane of E. coli, and it helps to maintain the cell envelope integrity of the E. coli cells at the lethal temperatures. Together, our data indicate that maintaining the cell envelope integrity is crucial for the E. coli cells to grow at high temperatures and also shed new light on the development of thermophilic bacteria for industrial application.T emperature is considered the most important single environmental factor that profoundly affects the structure and function of biomolecules. Each organism in nature has evolved to live at a certain optimal temperature range. Nonetheless, effective mechanisms have also been evolved for organisms to survive under nonoptimal temperature conditions, typically termed heat shock response (1, 2) and cold shock response (3). It is of great value to understand such mechanisms of living organisms for both unveiling the nature of life and exploring biotechnological application.Escherichia coli, being the most extensively studied bacterium and also a popularly utilized host cell for producing pharmaceutically important recombinant proteins, is known to be unable to grow at a temperature higher than 46.5°C (4-6). It has been widely reported that heterologous overexpression of certain exogenous molecular chaperones (7-11) or an endogenous transcriptional regulator (12) is able to significantly increase the viability of E. coli cells undergoing heat shock treatment at lethal temperatures (around 50°C). It is also well established that preincubating E. coli cells (13,14) and other organisms (reviewed in reference 15) at a sublethal temperature (e.g., 42°C) significantly increases the thermotolerance of the treated organisms at lethal temperatures. However, all these alternations usually would not allow the modified cells to permanently survive and even grow under such lethal temperatures. In two attempts at selecting heat-resistant phenotypes by using extensive experimental evolution, E. coli mutant strains that are able to grow at up to 48°C (16) or 48.5°C (5) were obtained, with growth at the latter temperature being partially related to the high level of expression of the molecular chaperone GroEL/GroES. In contrast, thermophilic bacteria have been found to grow effectively at optimal temperatures much higher than 50°C (17)(18)(19)(20). They are known...

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Dynamics of Escherichia coli at elevated temperatures: effect of temperature history and medium

Cited by 18 publications

References 56 publications

Experimental Evolution of a Facultative Thermophile from a Mesophilic Ancestor

Experimental Evolution of a Facultative Thermophile from a Mesophilic Ancestor

Over-expression and characterization of recombinant prefoldin from hyperthermophilic archaeum Pyrococcus furiosus in E. coli

A Small Heat Shock Protein Enables Escherichia coli To Grow at a Lethal Temperature of 50 C Conceivably by Maintaining Cell Envelope Integrity

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