%0 Journal Article
%A Della Corte, Dennis
%A van Beek, Hugo
%A Syberg, Falk
%A Schallmey, Marcus
%A Tobola, Felix
%A Cormann, Kai
%A Schlicker, Christine
%A Baumann, Philipp Tobias
%A Krumbach, Karin
%A Sokolowsky, Sascha
%A Morris, Connor J.
%A Grünberger, Alexander
%A Hofmann, Eckhard
%A Schröder, Gunnar F.
%A Marienhagen, Jan
%T Engineering and application of a biosensor with focused ligand specificity
%J Nature Communications
%V 11
%N 1
%@ 2041-1723
%C [London]
%I Nature Publishing Group UK
%M FZJ-2020-03061
%P 4851
%D 2020
%X Cell 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.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:32978386
%U <Go to ISI:>//WOS:000573735500001
%R 10.1038/s41467-020-18400-0
%U https://juser.fz-juelich.de/record/884040