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.
Highlights
Fluorine is a key element in the synthesis of molecules broadly used in medicine, agriculture and materials
A fluoride ion (F−)responsive riboswitch (FRS) has been described in bacteria and archaea, where it regulates the expression of genes involved in ion detoxification—typically controlling F− transporters[30]
We explored if a minimal FRS structure could be adapted as an orthogonal expression system in P. putida, which would afford the use of basal F− salts as inducers of gene expression
Summary
Fluorine is a key element in the synthesis of molecules broadly used in medicine, agriculture and materials. 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. The discovery of the fluorinase enzyme11,12 [5′-fluoro-5′-deoxyadenosine (5′-FDA) synthase] in Streptomyces and related Gram-positive bacterial species offered a unique opportunity to address this challenge This is the only enzyme known to incorporate inorganic fluoride (F−) into organic compounds by catalyzing the SN2 addition of F to the universal C1 donor S-adenosyl-L-methionine (SAM), thereby generating 5′-. Establishing de novo biofluorination in a robust, genetically tractable cell factory for cost-effective fluorometabolite biosynthesis, independent of feeding organofluorine or expensive precursors and inducers, is still a major challenge in metabolic engineering
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
