6xtv
From Proteopedia
FULL-LENGTH LTTR LYSG FROM CORYNEBACTERIUM GLUTAMICUM WITH BOUND EFFECTOR ARG
Structural highlights
FunctionLYSG_CORGL Positively regulates the expression of the exporter LysE. Induction requires the presence of a coinducer, which is either intracellular L-lysine, L-arginine or L-citrulline. L-histidine also acts as coinducer of lysE expression, but this amino acid is not exported by LysE (PubMed:11429454, PubMed:27350619). The lysEG system prevents bacteriostasis due to elevated L-lysine or L-arginine concentrations that arise during growth in the presence of peptides or in mutants possessing a deregulated biosynthesis pathway (PubMed:11429454).[1] [2] Publication Abstract from PubMedCell factories converting bio-based precursors to chemicals present an attractive avenue to a sustainable economy, yet screening of genetically diverse strain libraries to identify the best-performing whole-cell biocatalysts is a low-throughput endeavor. For this reason, transcriptional biosensors attract attention as they allow the screening of vast libraries when used in combination with fluorescence-activated cell sorting (FACS). However, broad ligand specificity of transcriptional regulators (TRs) often prohibits the development of such ultra-high-throughput screens. Here, we solve the structure of the TR LysG of Corynebacterium glutamicum, which detects all three basic amino acids. Based on this information, we follow a semi-rational engineering approach using a FACS-based screening/counterscreening strategy to generate an L-lysine insensitive LysG-based biosensor. This biosensor can be used to isolate L-histidine-producing strains by FACS, showing that TR engineering towards a more focused ligand spectrum can expand the scope of application of such metabolite sensors. Engineering and application of a biosensor with focused ligand specificity.,Della Corte D, van Beek HL, Syberg F, Schallmey M, Tobola F, Cormann KU, Schlicker C, Baumann PT, Krumbach K, Sokolowsky S, Morris CJ, Grunberger A, Hofmann E, Schroder GF, Marienhagen J Nat Commun. 2020 Sep 25;11(1):4851. doi: 10.1038/s41467-020-18400-0. PMID:32978386[3] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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