5B6I

Structure of fluorinase from Streptomyces sp. MA37

Summary for 5B6I

• Entry DOI: 10.2210/pdb5b6i/pdb
• Descriptor: Fluorinase, METHIONINE, ADENOSINE, ... (4 entities in total)
• Functional Keywords: transferase
• Biological source: Streptomyces sp. MA37
• Total number of polymer chains: 2
• Total formula weight: 69582.37

Authors

Xue, B.,Robinson, R.C. (deposition date: 2016-05-29, release date: 2016-10-26, Last modification date: 2023-11-08)

Primary citation

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.
Directed Evolution of a Fluorinase for Improved Fluorination Efficiency with a Non-native Substrate
Angew.Chem.Int.Ed.Engl., 55:14277-14280, 2016

PubMed 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'-[ 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 k value in the conversion of 5'-ClDA into S-adenosyl-l-methionine (SAM) but a reduced k value in the conversion of SAM into 5'-FDA.

PubMed: 27739177
DOI: 10.1002/anie.201606722

Experimental method

X-RAY DIFFRACTION (1.95 Å)