An Engineered E. coli Strain for Direct in Vivo Fluorination

Chembiochem. 2020 Jul 1;21(13):1856-1860. doi: 10.1002/cbic.202000051. Epub 2020 Mar 3.

Abstract

Selectively fluorinated compounds are found frequently in pharmaceutical and agrochemical products where currently 25-30 % of optimised compounds emerge from development containing at least one fluorine atom. There are many methods for the site-specific introduction of fluorine, but all are chemical and they often use environmentally challenging reagents. Biochemical processes for C-F bond formation are attractive, but they are extremely rare. In this work, the fluorinase enzyme, originally identified from the actinomycete bacterium Streptomyces cattleya, is engineered into Escherichia coli in such a manner that the organism is able to produce 5'-fluorodeoxyadenosine (5'-FDA) from S-adenosyl-l-methionine (SAM) and fluoride in live E. coli cells. Success required the introduction of a SAM transporter and deletion of the endogenous fluoride efflux capacity in order to generate an E. coli host that has the potential for future engineering of more elaborate fluorometabolites.

Keywords: Escherichia coli; SAM transporters; fluoride channels; fluorinases; halogenations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Deoxyadenosines / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Fluorine / chemistry
  • Fluorine / metabolism*
  • Genetic Engineering*
  • Halogenation
  • Isomerism
  • Oxidoreductases / genetics
  • Oxidoreductases / metabolism
  • S-Adenosylmethionine / metabolism
  • Streptomyces / enzymology

Substances

  • Bacterial Proteins
  • Deoxyadenosines
  • Fluorine
  • 2'-fluoro-2'-deoxyadenosine
  • S-Adenosylmethionine
  • Oxidoreductases
  • fluorinase

Supplementary concepts

  • Streptomyces cattleya