An Engineered E. coli Strain for Direct in Vivo Fluorination
Konstantinos Markakis1, Phillip T Lowe2, Liam Davison-Gates1
1Institute for Bioengineering, School of Engineering, University of Edinburgh, Faraday Building, King's Buildings, Colin Maclaurin Road, Edinburgh, EH9 3DW, UK.
Chembiochem : a European Journal of Chemical Biology
|February 1, 2020
Summary
Researchers engineered the fluorinase enzyme into E. coli, enabling live cells to produce 5'-fluorodeoxyadenosine (5'-FDA) from S-adenosyl-l-methionine (SAM) and fluoride. This breakthrough offers a sustainable biochemical route for creating fluorinated compounds.
Area of Science:
- Biochemistry
- Synthetic Biology
- Medicinal Chemistry
Background:
- Fluorinated compounds are crucial in pharmaceuticals and agrochemicals, with 25-30% of optimized compounds containing fluorine.
- Current methods for introducing fluorine are chemical, often using environmentally hazardous reagents.
- Biochemical C-F bond formation is highly desirable but exceptionally rare.
Purpose of the Study:
- To engineer a microbial host capable of producing fluorinated metabolites.
- To establish a sustainable, biocatalytic method for synthesizing 5 '-fluorodeoxyadenosine (5 '-FDA).
Main Methods:
- Engineered the fluorinase enzyme from Streptomyces cattleya into Escherichia coli.
- Introduced a S-adenosyl-l-methionine (SAM) transporter into E. coli.
- Deleted the endogenous fluoride efflux capacity in E. coli.
Main Results:
- Achieved successful production of 5 '-FDA from SAM and fluoride in live E. coli cells.
- Demonstrated the feasibility of using engineered E. coli as a biocatalyst for fluorination.
- Created an E. coli host suitable for future engineering of complex fluorometabolites.
Conclusions:
- The engineered E. coli strain provides a novel biocatalytic platform for producing 5 '-FDA.
- This work paves the way for developing sustainable biochemical methods for fluorinated compound synthesis.
- The developed E. coli host has potential for engineering more elaborate fluorinated metabolites.
