ArticleNature communications2022
CRISPR-assisted rational flux-tuning and arrayed CRISPRi screening of an L-proline exporter for L-proline hyperproduction.
Article in Nature communications, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed.
- N-terminal fusion length: The key to reliable and context-preserving regulatory sequence characterization.Synthetic and systems biotechnology · 2027Article
- Multi-omics analyses of evolvedSynthetic and systems biotechnology · 2026Article
- Systems metabolic engineering of Corynebacterium glutamicum for efficient production of L-isoleucine.Journal of animal science and biotechnology · 2026Article
- Harnessing the thermotolerant methylotroph Bacillus methanolicus for methanol-based synthetic L-proline production.Microbial cell factories · 2026Article
- Putative membrane protein Cgl2168 and its variant enhance thermotolerance inSynthetic and systems biotechnology · 2026Article
- Tailoring Corynebacterium glutamicum for Sustainable Biomanufacturing: From Traditional to Cutting-Edge Technologies.Molecular biotechnology · 2026Review
- Integrated transcriptomic and metabolomic analyses uncover novel genetic targets for enhanced l-tryptophan production inSynthetic and systems biotechnology · 2026Article
- Harnessing prokaryotic amino acid transporters for metabolic engineering: mechanisms and biotechnological applications.Synthetic and systems biotechnology · 2026Review
- Development of a CRISPR/Cas9 genome editing toolbox forSynthetic and systems biotechnology · 2025Article
- Engineered membraneless organelles inSynthetic and systems biotechnology · 2025Article
- Matrix regulation: a plug-and-tune method for combinatorial regulation in Saccharomyces cerevisiae.Nature communications · 2025Article
- Systems metabolic engineering ofSynthetic and systems biotechnology · 2025Article
- Engineering of L-threonine and L-proline biosensors by directed evolution of transcriptional regulator SerR and application for high-throughput screening.Bioresources and bioprocessing · 2025Article
- Microbial engineering for monocyclic aromatic compounds production.FEMS microbiology reviews · 2025Review
- CRISPR/Cas13X-assisted programmable and multiplexed translation regulation for controlled biosynthesis.Nucleic acids research · 2025Article
- Automated characterization and analysis of expression compatibility between regulatory sequences and metabolic genes inSynthetic and systems biotechnology · 2024Article
- A previously unidentified sugar transporter for engineering of high-yield Streptomyces.Applied microbiology and biotechnology · 2024Article
- CRISPRi-mediated metabolic switch enables concurrent aerobic and synthetic anaerobic fermentations in engineered consortium.Nature communications · 2024Article
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- An Ultrasensitive Biosensor for Probing Subcellular Distribution and Mitochondrial Transport of l-2-Hydroxyglutarate.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
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Authors and funding
16 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Development of hyperproducing strains is important for biomanufacturing of biochemicals and biofuels but requires extensive efforts to engineer cellular metabolism and discover functional components. Herein, we optimize and use the CRISPR-assisted editing and CRISPRi screening methods to convert a wild-type Corynebacterium glutamicum to a hyperproducer of L-proline, an amino acid with medicine, feed, and food applications. To facilitate L-proline production, feedback-deregulated variants of key biosynthetic enzyme γ-glutamyl kinase are screened using CRISPR-assisted single-stranded DNA recombineering. To increase the carbon flux towards L-proline biosynthesis, flux-control genes predicted by in silico analysis are fine-tuned using tailored promoter libraries. Finally, an arrayed CRISPRi library targeting all 397 transporters is constructed to discover an L-proline exporter Cgl2622. The final plasmid-, antibiotic-, and inducer-free strain produces L-proline at the level of 142.4 g/L, 2.90 g/L/h, and 0.31 g/g. The CRISPR-assisted strain development strategy can be used for engineering industrial-strength strains for efficient biomanufacturing.
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