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000884040 1001_ $$0P:(DE-Juel1)145868$$aDella Corte, Dennis$$b0
000884040 245__ $$aEngineering and application of a biosensor with focused ligand specificity
000884040 260__ $$a[London]$$bNature Publishing Group UK$$c2020
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000884040 520__ $$aCell 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.
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000884040 7001_ $$0P:(DE-Juel1)167342$$avan Beek, Hugo$$b1
000884040 7001_ $$0P:(DE-HGF)0$$aSyberg, Falk$$b2
000884040 7001_ $$0P:(DE-Juel1)136701$$aSchallmey, Marcus$$b3
000884040 7001_ $$0P:(DE-Juel1)157947$$aTobola, Felix$$b4
000884040 7001_ $$0P:(DE-Juel1)167285$$aCormann, Kai$$b5
000884040 7001_ $$0P:(DE-HGF)0$$aSchlicker, Christine$$b6
000884040 7001_ $$0P:(DE-Juel1)174462$$aBaumann, Philipp Tobias$$b7$$ufzj
000884040 7001_ $$0P:(DE-Juel1)128973$$aKrumbach, Karin$$b8$$ufzj
000884040 7001_ $$0P:(DE-Juel1)138744$$aSokolowsky, Sascha$$b9$$ufzj
000884040 7001_ $$0P:(DE-HGF)0$$aMorris, Connor J.$$b10
000884040 7001_ $$0P:(DE-Juel1)143612$$aGrünberger, Alexander$$b11
000884040 7001_ $$0P:(DE-HGF)0$$aHofmann, Eckhard$$b12
000884040 7001_ $$0P:(DE-Juel1)132018$$aSchröder, Gunnar F.$$b13
000884040 7001_ $$0P:(DE-Juel1)144031$$aMarienhagen, Jan$$b14$$eCorresponding author$$ufzj
000884040 773__ $$0PERI:(DE-600)2553671-0$$a10.1038/s41467-020-18400-0$$gVol. 11, no. 1, p. 4851$$n1$$p4851$$tNature Communications$$v11$$x2041-1723$$y2020
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