Sun, H., Yeo, W. L., Lim, Y. H., Chew, X., Smith, D. J., Xue, B., … 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. doi:10.1002/anie.201606722
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'-[18 F]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 kcat value in the conversion of 5'-ClDA into S-adenosyl-l-methionine (SAM) but a reduced kcat value in the conversion of SAM into 5'-FDA.
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Publisher Copyright
Funding Info:
This work was funded by the GlaxoSmithKline - Singapore Economic Development Board partnership for green and sustainable manufacturing, A*STAR ICES, and A*STAR IMCB.
Description:
This is the peer reviewed version of the following article: Sun, H., Yeo, W. L., Lim, Y. H., Chew, X., Smith, D. J., Xue, B., … 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. doi:10.1002/anie.201606722, which has been published in final form at http://dx.doi.org/10.1002/anie.201606722. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.