Pyruvate-responsive genetic circuits for dynamic control of central metabolism

Xu, Xianhao; Li, Xueliang; Liu, Yanfeng; Zhu, Yangyang; Li, Jianghua; Du, Guocheng; Chen, Jian; Ledesma-Amaro, Rodrigo; Liu, Long

doi:10.1038/s41589-020-0637-3

Cited by 112 publications

(73 citation statements)

References 52 publications

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“…ynthetic biology aiming to perform computational programs in living cells enables revolutionary developments in biotechnology 1,2 . Notably, scalable and robust gene circuits for uni-and bidirectional control of target genes over various timings and levels triggered by specific signals are desirable for diverse applications [3][4][5] . Generally, a dynamic control system of customized functions can be achieved by leveraging a variety of signal-sensing modules, including chemicals 6,7 , intermediate metabolites [8][9][10] , temperature [11][12][13] , light [14][15][16] , and cell population 17 responsive biosensors.…”

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confidence: 99%

Reversible thermal regulation for bifunctional dynamic control of gene expression in Escherichia coli

et al. 2021

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Genetically programmed circuits allowing bifunctional dynamic regulation of enzyme expression have far-reaching significances for various bio-manufactural purposes. However, building a bio-switch with a post log-phase response and reversibility during scale-up bioprocesses is still a challenge in metabolic engineering due to the lack of robustness. Here, we report a robust thermosensitive bio-switch that enables stringent bidirectional control of gene expression over time and levels in living cells. Based on the bio-switch, we obtain tree ring-like colonies with spatially distributed patterns and transformer cells shifting among spherical-, rod- and fiber-shapes of the engineered Escherichia coli. Moreover, fed-batch fermentations of recombinant E. coli are conducted to obtain ordered assembly of tailor-made biopolymers polyhydroxyalkanoates including diblock- and random-copolymer, composed of 3-hydroxybutyrate and 4-hydroxybutyrate with controllable monomer molar fraction. This study demonstrates the possibility of well-organized, chemosynthesis-like block polymerization on a molecular scale by reprogrammed microbes, exemplifying the versatility of thermo-response control for various practical uses.

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confidence: 99%

Reversible thermal regulation for bifunctional dynamic control of gene expression in Escherichia coli

et al. 2021

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“…Biosensor-assisted cell selection has been proven to be an effective approach to improve the quality of the microbial population and enhance the microbial biosynthesis performance [ 16 , 18 , 21 , 27 – 29 ]. For this study, this approach was adopted to engineer the upstream co-culture strain.…”

Section: Resultsmentioning

confidence: 99%

Development and optimization of a microbial co-culture system for heterologous indigo biosynthesis

et al. 2021

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Background Indigo is a color molecule with a long history of being used as a textile dye. The conventional production methods are facing increasing economy, sustainability and environmental challenges. Therefore, developing a green synthesis method converting renewable feedstocks to indigo using engineered microbes is of great research and application interest. However, the efficiency of the indigo microbial biosynthesis is still low and needs to be improved by proper metabolic engineering strategies. Results In the present study, we adopted several metabolic engineering strategies to establish an efficient microbial biosynthesis system for converting renewable carbon substrates to indigo. First, a microbial co-culture was developed using two individually engineered E. coli strains to accommodate the indigo biosynthesis pathway, and the balancing of the overall pathway was achieved by manipulating the ratio of co-culture strains harboring different pathway modules. Through carbon source optimization and application of biosensor-assisted cell selection circuit, the indigo production was improved significantly. In addition, the global transcription machinery engineering (gTME) approach was utilized to establish a high-performance co-culture variant to further enhance the indigo production. Through the step-wise modification of the established system, the indigo bioproduction reached 104.3 mg/L, which was 11.4-fold higher than the parental indigo producing strain. Conclusion This work combines modular co-culture engineering, biosensing, and gTME for addressing the challenges of the indigo biosynthesis, which has not been explored before. The findings of this study confirm the effectiveness of the developed approach and offer a new perspective for efficient indigo bioproduction. More broadly, this innovative approach has the potential for wider application in future studies of other valuable biochemicals’ biosynthesis.

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“…Secondly, assembling catalytic enzymes to rewire the endogenous flux towards metabolic targets is commonly employed to achieve prototype success of target products from 0 to 1, namely pathway construction. Further optimization leveraging static regulation, mainly refers to bypass deletion and flux tuning, and dynamic regulation [96] allowing gene expression control over time and levels, is a proven strategy to generate significant breakthrough from 1 to 100. Thirdly, a rigorous scale-up test including medium and feeding strategy optimization can obtain iterative bioprocess of refinement.…”

Section: Metabolic Engineering In Halomonas Sppmentioning

confidence: 99%

Halomonasas a chassis

Chen

2021

Essays in Biochemistry

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With the rapid development of systems and synthetic biology, the non-model bacteria, Halomonas spp., have been developed recently to become a cost-competitive platform for producing a variety of products including polyesters, chemicals and proteins owing to their contamination resistance and ability of high cell density growth at alkaline pH and high salt concentration. These salt-loving microbes can partially solve the challenges of current industrial biotechnology (CIB) which requires high energy-consuming sterilization to prevent contamination as CIB is based on traditional chassis, typically, Escherichia coli, Bacillus subtilis, Pseudomonas putida and Corynebacterium glutamicum. The advantages and current status of Halomonas spp. including their molecular biology and metabolic engineering approaches as well as their applications are reviewed here. Moreover, a systematic strain engineering streamline, including product-based host development, genetic parts mining, static and dynamic optimization of modularized pathways and bioprocess-inspired cell engineering are summarized. All of these developments result in the term called next-generation industrial biotechnology (NGIB). Increasing efforts are made to develop their versatile cell factories powered by synthetic biology to demonstrate a new biomanufacturing strategy under open and continuous processes with significant cost-reduction on process complexity, energy, substrates and fresh water consumption.

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Pyruvate-responsive genetic circuits for dynamic control of central metabolism

Cited by 112 publications

References 52 publications

Reversible thermal regulation for bifunctional dynamic control of gene expression in Escherichia coli

Reversible thermal regulation for bifunctional dynamic control of gene expression in Escherichia coli

Development and optimization of a microbial co-culture system for heterologous indigo biosynthesis

Halomonasas a chassis

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