Laboratory-Evolved Mutants of an Exogenous Global Regulator, IrrE from Deinococcus radiodurans, Enhance Stress Tolerances of Escherichia coli
Open accessAbstract
BACKGROUND: The tolerance of cells toward different stresses is very important for industrial strains of microbes, but difficult to improve by the manipulation of single genes. Traditional methods for enhancing cellular tolerances are inefficient and time-consuming. Recently, approaches employing global transcriptional or translational engineering methods have been increasingly explored. We found that an exogenous global regulator, irrE from an extremely radiation-resistant bacterium, Deinococcus radiodurans, has the potential to act as a global regulator in Escherichia coli, and that laboratory-evolution might be applied to alter this regulator to elicit different phenotypes for E. coli. METHODOLOGY/PRINCIPAL FINDINGS: To extend the methodology for strain improvement and to obtain higher tolerances toward different stresses, we here describe an approach of engineering irrE gene in E. coli. An irrE library was constructed by randomly mutating the gene, and this library was then selected for tolerance to ethanol, butanol and acetate stresses. Several mutants showing significant tolerances were obtained and characterized. The tolerances of E. coli cells containing these mutants were enhanced 2 to 50-fold, based on cell growth tests using different concentrations of alcohols or acetate, and enhanced 10 to 100-fold based on ethanol or butanol shock experiments. Intracellular reactive oxygen species (ROS) assays showed that intracellular ROS levels were sharply reduced for cells containing the irrE mutants. Sequence analysis of the mutants revealed that the mutations distribute cross all three domains of the protein. CONCLUSIONS: To our knowledge, this is the first time that an exogenous global regulator has been artificially evolved to suit its new host. The successes suggest the possibility of improving tolerances of industrial strains by introducing and engineering exogenous global regulators, such as those from extremophiles. This new approach can be applied alone or in combination with other global methods, such as global transcriptional machinery engineering (gTME) for strain improvements.
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The SPANC balance in E. coli is governed by strict control of processes such as stress and stringent response.
“This SPANC balance was intensively studied in E. coli and is governed by strict control of processes such as stress and stringent response [158,159].”
From Adaptive laboratory evolution—Principles and applications for biotechnology (2013) · cites this paper as [159]
Microbial cells have an intrinsic capacity to balance self preservation (stress protection) and nutritional competence (SPANC).
“Microbial cells have an intrinsic capacity to balance self preservation (stress protection) and nutritional competence (SPANC).”
From Adaptive laboratory evolution—Principles and applications for biotechnology (2013) · cites this paper as [159]
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- Authors
- Chen, T., Wang, J., Yang, R., Li, J., Lin, M., & Lin, Z.
- Affiliations
- Tsinghua University (CN)
- Chinese Academy of Agricultural Sciences (CN)
- Corresponding author
- Lin, Min
- Lin, Zhanglin
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2011-01-18
- Volume
- 6
- Issue
- 1
- Pages
- e16228
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC3022760
- OpenAlex
- W2066542358
- Cited by
- 81
- References
- 46
- Citations by year
- 2026: 2 · 2024: 8 · 2023: 2 · 2022: 3 · 2021: 5 · 2020: 9 · 2019: 3 · 2018: 5 · 2017: 4 · 2016: 1 · 2015: 8 · 2014: 10 · 2013: 12 · 2012: 8
- FWCI
- 4.02
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- Deinococcus radiodurans, global regulators, Escherichia coli, experimental evolution, stress tolerance, ethanol tolerance, butanol tolerance, acetate tolerance, reactive oxygen species, extremophile proteins, industrial strain improvement
- MeSH terms
- Escherichia coli, Industrial Microbiology, Mutation, Stress, Physiological, Gene Library, Reactive Oxygen Species, Directed Molecular Evolution, Deinococcus, Regulatory Elements, Transcriptional
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