Directed Evolution of a Fluorinase for Improved Fluorination Efficiency with a Non-native Substrate

Huihua Sun, Wan Lin Yeo, Yee Hwee Lim, Xinying Chew, Derek John Smith, Bo Xue, Kok Ping Chan, Robert C. Robinson, Edward G. Robins, Huimin Zhao, Ee Lui Ang

Research output: Contribution to journalArticle

Abstract

Fluorinases offer an environmentally friendly alternative for selective fluorination under mild conditions. However, their diversity is limited in nature and they have yet to be engineered through directed evolution. Herein, we report the directed evolution of the fluorinase FlA1 for improved conversion of the non-native substrate 5′-chloro-5′-deoxyadenosine (5′-ClDA) into 5′-fluoro-5′-deoxyadenosine (5′-FDA). The evolved variants, fah2081 (A279Y) and fah2114 (F213Y, A279L), were successfully applied in the radiosynthesis of 5′-[18F]FDA, with overall radiochemical conversion (RCC) more than 3-fold higher than wild-type FlA1. Kinetic studies of the two-step reaction revealed that the variants show a significantly improved kcatvalue in the conversion of 5′-ClDA into S-adenosyl-l-methionine (SAM) but a reduced kcatvalue in the conversion of SAM into 5′-FDA.

Original languageEnglish (US)
Pages (from-to)14277-14280
Number of pages4
JournalAngewandte Chemie - International Edition
Volume55
Issue number46
DOIs
StatePublished - Jan 1 2016

Keywords

  • biocatalysis
  • directed evolution
  • fluorinases
  • halogenation
  • radiosynthesis

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

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  • Cite this

    Sun, H., Yeo, W. L., Lim, Y. H., Chew, X., Smith, D. J., Xue, B., Chan, K. P., Robinson, R. C., Robins, E. G., Zhao, H., & Ang, E. L. (2016). Directed Evolution of a Fluorinase for Improved Fluorination Efficiency with a Non-native Substrate. Angewandte Chemie - International Edition, 55(46), 14277-14280. https://doi.org/10.1002/anie.201606722