Journal Article

FZJ-2020-03061

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Engineering and application of a biosensor with focused ligand specificity

Della Corte, D.FZJ* ; van Beek, H.FZJ* ; Syberg, F. ; Schallmey, M.FZJ* ; Tobola, F.FZJ* ; Cormann, K.FZJ* ; Schlicker, C. ; Baumann, P. T.FZJ* ; Krumbach, K.FZJ* ; Sokolowsky, S.FZJ* ; Morris, C. J. ; Grünberger, A.FZJ* ; Hofmann, E. ; Schröder, G. F.FZJ* ; Marienhagen, J. (Corresponding author)FZJ*

2020
Nature Publishing Group UK [London]

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Please use a persistent id in citations: http://hdl.handle.net/2128/25800  doi:10.1038/s41467-020-18400-0

Abstract: 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.

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Contributing Institute(s):

1. Biotechnologie (IBG-1)
2. Strukturbiochemie (IBI-7)

Research Program(s):

1. 581 - Biotechnology (POF3-581) (POF3-581)

Appears in the scientific report 2020

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