TY  - JOUR
AU  - Della Corte, Dennis
AU  - van Beek, Hugo
AU  - Syberg, Falk
AU  - Schallmey, Marcus
AU  - Tobola, Felix
AU  - Cormann, Kai
AU  - Schlicker, Christine
AU  - Baumann, Philipp Tobias
AU  - Krumbach, Karin
AU  - Sokolowsky, Sascha
AU  - Morris, Connor J.
AU  - Grünberger, Alexander
AU  - Hofmann, Eckhard
AU  - Schröder, Gunnar F.
AU  - Marienhagen, Jan
TI  - Engineering and application of a biosensor with focused ligand specificity
JO  - Nature Communications
VL  - 11
IS  - 1
SN  - 2041-1723
CY  - [London]
PB  - Nature Publishing Group UK
M1  - FZJ-2020-03061
SP  - 4851
PY  - 2020
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:32978386
UR  - <Go to ISI:>//WOS:000573735500001
DO  - DOI:10.1038/s41467-020-18400-0
UR  - https://juser.fz-juelich.de/record/884040
ER  -