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A fluoride-responsive genetic circuit enables in vivo biofluorination in engineered Pseudomonas putida

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Calero_2020_NC_Fluoride_responsive_CC.pdf (911.7Kb)
Date
07/10/2020
Author
Calero, Patricia
Volke, Daniel C.
Lowe, Phillip
Gotfredsen, Charlotte H.
O'Hagan, David
Nikel, Pablo I.
Funder
European Commission
Grant ID
814418
Keywords
QD Chemistry
QR Microbiology
DAS
BDC
R2C
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Abstract
Fluorine is a key element in the synthesis of molecules broadly used in medicine, agriculture and materials. Addition of fluorine to organic structures represents a unique strategy for tuning molecular properties, yet this atom is rarely found in Nature and approaches to integrate fluorometabolites into the biochemistry of living cells are scarce. In this work, synthetic gene circuits for organofluorine biosynthesis are implemented in the platform bacterium Pseudomonas putida. By harnessing fluoride-responsive riboswitches and the orthogonal T7 RNA polymerase, biochemical reactions needed for in vivo biofluorination are wired to the presence of fluoride (i.e. circumventing the need of feeding expensive additives). Biosynthesis of fluoronucleotides and fluorosugars in engineered P. putida is demonstrated with mineral fluoride both as only fluorine source (i.e. substrate of the pathway) and as inducer of the synthetic circuit. This approach expands the chemical landscape of cell factories by providing alternative biosynthetic strategies towards fluorinated building-blocks.
Citation
Calero , P , Volke , D C , Lowe , P , Gotfredsen , C H , O'Hagan , D & Nikel , P I 2020 , ' A fluoride-responsive genetic circuit enables in vivo biofluorination in engineered Pseudomonas putida ' , Nature Communications , vol. 11 , 5045 . https://doi.org/10.1038/s41467-020-18813-x
Publication
Nature Communications
Status
Peer reviewed
DOI
https://doi.org/10.1038/s41467-020-18813-x
ISSN
2041-1723
Type
Journal article
Rights
Copyright © 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/.
Description
Funding: This work was supported by grants from The Novo Nordisk Foundation (NNF10CC1016517 and NNF18OC0034818), the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 814418 (SinFonia) and the Danish Council for Independent Research (SWEET, DFF-Research Project 8021-00039B) to P.I.N. We also thank the NMR Center at the Technical University of Denmark and the Villum Foundation for facilitating access to the 600 and 800 MHz NMR spectrometers.
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  • University of St Andrews Research
URI
http://hdl.handle.net/10023/20765

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