Biosensor-based isolation of amino acid-producing Vibrio natriegens strains
| Autor: | Sophia Lorke, Johanna Wiechert, Julia Frunzke, Jan Marienhagen, Philipp T. Baumann, Roberto Giuseppe Stella, Felix Münstermann, Astrid Wirtz |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Transcription factor-based biosensors
QH301-705.5 Endocrinology Diabetes and Metabolism High-throughput screening Biomedical Engineering Mutagenesis (molecular biology technique) Vibrio natriegens Corynebacterium glutamicum Metabolic engineering 03 medical and health sciences Full Length Article ddc:610 Biology (General) 030304 developmental biology chemistry.chemical_classification 0303 health sciences biology 030306 microbiology Effector Amino acid production biology.organism_classification 3. Good health Amino acid Biochemistry chemistry TP248.13-248.65 Bacteria Biotechnology |
| Zdroj: | Metabolic engineering communications 13, e00187-(2021). doi:10.1016/j.mec.2021.e00187 Metabolic engineering communications 13, e00187 (2021). doi:10.1016/j.mec.2021.e00187 Metabolic Engineering Communications, Vol 13, Iss, Pp e00187-(2021) Metabolic Engineering Communications |
| DOI: | 10.1016/j.mec.2021.e00187 |
| Popis: | The marine bacterium Vibrio natriegens has recently been demonstrated to be a promising new host for molecular biology and next generation bioprocesses. V. natriegens is a Gram-negative, non-pathogenic slight-halophilic bacterium, with a high nutrient versatility and a reported doubling time of under 10 min. However, V. natriegens is not an established model organism yet, and further research is required to promote its transformation into a microbial workhorse. In this work, the potential of V. natriegens as an amino acid producer was investigated. First, the transcription factor-based biosensor LysG, from Corynebacterium glutamicum, was adapted for expression in V. natriegens to facilitate the detection of positively charged amino acids. A set of different biosensor variants were constructed and characterized, using the expression of a fluorescent protein as sensor output. After random mutagenesis, one of the LysG-based sensors was used to screen for amino acid producer strains. Here, fluorescence-activated cell sorting enabled the selective sorting of highly fluorescent cells, i.e. potential producer cells. Using this approach, individual L-lysine, L-arginine and L-histidine producers could be obtained producing up to 1 mM of the effector amino acid, extracellularly. Genome sequencing of the producer strains provided insight into the amino acid production metabolism of V. natriegens. This work demonstrates the successful expression and application of transcription factor-based biosensors in V. natriegens and provides insight into the underlying physiology, forming a solid basis for further development of this promising microbe. Highlights • Vibrio natriegens is a promising new host for biotechnology. • Transcription factor-based biosensors were expressed in V. natriegens. • Mutagenesis and screening using FACS provided amino acid producing mutants. • Genome sequencing revealed several causal mutations leading to amino acid production. • These results will support further efforts to develop V. natriegens as a production host. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|