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Engineering and application of a biosensor with focused ligand specificity
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 a...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7519686/ https://www.ncbi.nlm.nih.gov/pubmed/32978386 http://dx.doi.org/10.1038/s41467-020-18400-0 |
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author | Della Corte, Dennis van Beek, Hugo L. Syberg, Falk Schallmey, Marcus Tobola, Felix Cormann, Kai U. Schlicker, Christine Baumann, Philipp T. Krumbach, Karin Sokolowsky, Sascha Morris, Connor J. Grünberger, Alexander Hofmann, Eckhard Schröder, Gunnar F. Marienhagen, Jan |
author_facet | Della Corte, Dennis van Beek, Hugo L. Syberg, Falk Schallmey, Marcus Tobola, Felix Cormann, Kai U. Schlicker, Christine Baumann, Philipp T. Krumbach, Karin Sokolowsky, Sascha Morris, Connor J. Grünberger, Alexander Hofmann, Eckhard Schröder, Gunnar F. Marienhagen, Jan |
author_sort | Della Corte, Dennis |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7519686 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-75196862020-10-14 Engineering and application of a biosensor with focused ligand specificity Della Corte, Dennis van Beek, Hugo L. Syberg, Falk Schallmey, Marcus Tobola, Felix Cormann, Kai U. Schlicker, Christine Baumann, Philipp T. Krumbach, Karin Sokolowsky, Sascha Morris, Connor J. Grünberger, Alexander Hofmann, Eckhard Schröder, Gunnar F. Marienhagen, Jan Nat Commun Article 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. Nature Publishing Group UK 2020-09-25 /pmc/articles/PMC7519686/ /pubmed/32978386 http://dx.doi.org/10.1038/s41467-020-18400-0 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Della Corte, Dennis van Beek, Hugo L. Syberg, Falk Schallmey, Marcus Tobola, Felix Cormann, Kai U. Schlicker, Christine Baumann, Philipp T. Krumbach, Karin Sokolowsky, Sascha Morris, Connor J. Grünberger, Alexander Hofmann, Eckhard Schröder, Gunnar F. Marienhagen, Jan Engineering and application of a biosensor with focused ligand specificity |
title | Engineering and application of a biosensor with focused ligand specificity |
title_full | Engineering and application of a biosensor with focused ligand specificity |
title_fullStr | Engineering and application of a biosensor with focused ligand specificity |
title_full_unstemmed | Engineering and application of a biosensor with focused ligand specificity |
title_short | Engineering and application of a biosensor with focused ligand specificity |
title_sort | engineering and application of a biosensor with focused ligand specificity |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7519686/ https://www.ncbi.nlm.nih.gov/pubmed/32978386 http://dx.doi.org/10.1038/s41467-020-18400-0 |
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