Engineering a riboswitch-based genetic platform for the self-directed evolution of acid-tolerant phenotypes

Pham, Hoang Long; Wong, Adison; Chua, Niying; Teo, Wei Suong; Yew, Wen Shan; Chang, Matthew Wook

doi:10.1038/s41467-017-00511-w

Cited by 82 publications

(62 citation statements)

References 53 publications

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“…Several studies have employed combinatory libraries of regulatory elements for optimizing the heterologous expression of biosynthetic pathways 32–34 , and different molecular strategies haven been used for the rational improvement of bacterial robustness under different conditions, such as temperature, pH and ethanol concentration 35–38 . However, to the best of our knowledge, this is the first time such a combinatory approach was taken to generate a library of truly synthetic tolerance clusters composed by seemingly unrelated resistance genes.…”

Section: Resultsmentioning

confidence: 99%

Turning the screw: engineering extreme pH resistance inEscherichia colithrough combinatorial synthetic operons

Siqueira¹,

Silva‐Rocha²,

Guazzaroni³

2020

Preprint

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Adoption of microorganisms as platforms for sustainable biobased production requires host cells to be able to withstand harsh industrial conditions, which are usually far from the ones where these organisms are naturally adapted to thrive. However, novel survival mechanisms unearthed by the study of microbiomes from extreme habitats may be exploited to enhance microbial robustness under the strict conditions needed for different applications. In this work, synthetic biology approaches were used to engineer enhanced acidic tolerance in Escherichia coli under extreme conditions through the characterization of a library of twenty-seven unique operons composed of combinatorial assemblies of three novel genes from an extreme environment and three synthetic ribosome binding sites. The results here presented illustrate the efficacy of combining different metagenomic genes for tolerance in truly synthetic genetic operons, as expression of these gene clusters increased hundred-fold the survival percentage of cells exposed to an acidic shock in minimal media at pH 1.9 under aerobic conditions.

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Section: Resultsmentioning

confidence: 99%

Turning the screw: engineering extreme pH resistance inEscherichia colithrough combinatorial synthetic operons

Siqueira¹,

Silva‐Rocha²,

Guazzaroni³

2020

Preprint

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“…This dynamic system has been used to increase the production of l ‐tyrosine and isoprenoids in engineered E. coli strains. Similarly, Pham et al designed a riboswitch‐based pH‐sensing genetic device to control gene expression according to the environmental pH and used this device for evolutionary engineering of E. coli for improved tolerance to a broad spectrum of organic acids.…”

Section: Strategies To Increase Phenotypic Variation In Bacterial Popmentioning

confidence: 99%

Evolutionary Approaches for Engineering Industrially Relevant Phenotypes in Bacterial Cell Factories

Fernández-Cabezón

Cros

Nikel

2019

Biotechnology Journal

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The bio‐based production of added‐value compounds (with applications as pharmaceuticals, biofuels, food ingredients, and building blocks) using bacterial platforms is a well‐established industrial activity. The design and construction of microbial cell factories (MCFs) with robust and stable industrially relevant phenotypes, however, remains one of the biggest challenges of contemporary biotechnology. In this review, traditional and cutting‐edge approaches for optimizing the performance of MCFs for industrial bioprocesses, rooted on the engineering principle of natural evolution (i.e., genetic variation and selection), are discussed. State‐of‐the‐art techniques to manipulate and increase genetic variation in bacterial populations and to construct combinatorial libraries of strains, both globally (i.e., genome level) and locally (i.e., individual genes or pathways, and entire sections and gene clusters of the bacterial genome) are presented. Cutting‐edge screening and selection technologies applied to isolate MCFs displaying enhanced phenotypes are likewise discussed. The review article is closed by presenting future trends in the design and construction of a new generation of MCFs that will contribute to the long‐sought‐after transformation from a petrochemical industry to a veritable sustainable bio‐based industry.

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“…If the taRNA is transcribed, it binds the crRNA and hence releases the RBS to allow translation (Figure F) . Given the success of riboswitches in cyanobacteria and high portability of riboswitches between microorganisms, they hold promise for strain engineering and pathway optimization, as demonstrated in conventional hosts …”

Section: Programmable Regulationsmentioning

confidence: 99%

Synthetic Biology Toolkits for Metabolic Engineering of Cyanobacteria

Xia

Ling

Foo

et al. 2019

Biotechnology Journal

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Cyanobacteria are of great importance to Earth's ecology. Due to their capability in photosynthesis and C1 metabolism, they are ideal microbial chassis that can be engineered for direct conversion of carbon dioxide and solar energy into biofuels and biochemicals. Facilitated by the elucidation of the basic biology of the photoautotrophic microbes and rapid advances in synthetic biology, genetic toolkits have been developed to enable implementation of nonnatural functionalities in engineered cyanobacteria. Hence, cyanobacteria are fast becoming an emerging platform in synthetic biology and metabolic engineering. Herein, the progress made in the synthetic biology toolkits for cyanobacteria and their utilization for transforming cyanobacteria into microbial cell factories for sustainable production of biofuels and biochemicals is outlined. Current techniques in heterologous gene expression, strategies in genome editing, and development of programmable regulatory parts and modules for engineering cyanobacteria towards biochemical production are discussed and prospected. As cyanobacteria synthetic biology is still in its infancy, apart from the achievements made, the difficulties and challenges in applying and developing genetic toolkits in cyanobacteria for biochemical production are also evaluated.

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Engineering a riboswitch-based genetic platform for the self-directed evolution of acid-tolerant phenotypes

Cited by 82 publications

References 53 publications

Turning the screw: engineering extreme pH resistance inEscherichia colithrough combinatorial synthetic operons

Turning the screw: engineering extreme pH resistance inEscherichia colithrough combinatorial synthetic operons

Evolutionary Approaches for Engineering Industrially Relevant Phenotypes in Bacterial Cell Factories

Synthetic Biology Toolkits for Metabolic Engineering of Cyanobacteria

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