ArticleNucleic acids research2013
Recombineering in Corynebacterium glutamicum combined with optical nanosensors: a general strategy for fast producer strain generation.
Article in Nucleic acids research, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 57 papers.
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Who cites it
57 citing papers in PubMed, 154 citations in OpenAlex.
- Performance comparison of downstream unit operations and process chains for the purification of amino acids for food, feed and pharma applications.Biotechnology reports (Amsterdam, Netherlands) · 2026Review
- Tailoring Corynebacterium glutamicum for Sustainable Biomanufacturing: From Traditional to Cutting-Edge Technologies.Molecular biotechnology · 2026Review
- Advances in gene editing tools for four typical Gram-positive bacteria.Frontiers in microbiology · 2026Review
- Development of a CRISPR/Cas9 genome editing toolbox forSynthetic and systems biotechnology · 2025Article
- Recent advances in the Design-Build-Test-Learn (DBTL) cycle for systems metabolic engineering of Corynebacterium glutamicum.Journal of microbiology (Seoul, Korea) · 2025Review
- High Throughput Screening of Transcription Factor LysG for Constructing a Better Lysine Biosensor.Biosensors · 2024Article
- Expanding the CRISPR Toolbox for Engineering Lycopene Biosynthesis inMicroorganisms · 2024Article
- Rapid combinatorial rewiring of metabolic networks for enhanced poly(3-hydroxybutyrate) production in Corynebacterium glutamicum.Microbial cell factories · 2023Article
- Synthetic biology tools for engineeringComputational and structural biotechnology journal · 2023Review
- Recombineering in Non-Model Bacteria.Current protocols · 2022Article
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- Metabolic Footprinting of Microbial Systems Based on Comprehensive In Silico Predictions of MS/MS Relevant Data.Metabolites · 2022Article
- CRISPR-assisted rational flux-tuning and arrayed CRISPRi screening of an L-proline exporter for L-proline hyperproduction.Nature communications · 2022Article
- CRISPR/Cas tools for enhancing the biopreservation ability of lactic acid bacteria in aquatic products.Frontiers in bioengineering and biotechnology · 2022Review
- Transcriptional Regulation of the Creatine Utilization Genes ofFrontiers in bioengineering and biotechnology · 2022Article
- ReScribe: An Unrestrained Tool Combining Multiplex Recombineering and Minimal-PAM ScCas9 for Genome RecodingACS synthetic biology · 2021Article
- Identification of phage recombinase function unit in genus Corynebacterium.Applied microbiology and biotechnology · 2021Article
- Programming adaptive laboratory evolution of 4-hydroxyisoleucine production driven by a lysine biosensor in Corynebacterium glutamicum.AMB Express · 2021Article
- Need for speed: evaluation of dilute and shoot-mass spectrometry for accelerated metabolic phenotyping in bioprocess development.Analytical and bioanalytical chemistry · 2021Article
- Advances and Perspectives for Genome Editing Tools ofFrontiers in microbiology · 2021Review
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recombineering in bacteria is a powerful technique for genome reconstruction, but until now, it was not generally applicable for development of small-molecule producers because of the inconspicuous phenotype of most compounds of biotechnological relevance. Here, we establish recombineering for Corynebacterium glutamicum using RecT of prophage Rac and combine this with our recently developed nanosensor technology, which enables the detection and isolation of productive mutants at the single-cell level via fluorescence-activated cell sorting (FACS). We call this new technology RecFACS, which we use for genomic site-directed saturation mutagenesis without relying on pre-constructed libraries to directly isolate L-lysine-producing cells. A mixture of 19 different oligonucleotides was used targeting codon 81 in murE of the wild-type, at a locus where one single mutation is known to cause L-lysine production. Using RecFACS, productive mutants were screened and isolated. Sequencing revealed 12 different amino acid exchanges in the targeted murE codon, which caused different L-lysine production titers. Apart from introducing a rapid genome construction technology for C. glutamicum, the present work demonstrates that RecFACS is suitable to simply create producers as well as genetic diversity in one single step, thus establishing a new general concept in synthetic biology.
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