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Lookup NU author(s): Dr Jon Marles-WrightORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© The Royal Society of Chemistry.Enantiopure amines are key building blocks in the synthesis of many pharmaceuticals, so a route to their production is a current goal for biocatalysis. The stereo-inverting d-phenylglycine aminotransferase (d-PhgAT), isolated from Pseudomonas stutzeri ST-201, catalyses the reversible transamination from l-glutamic acid to benzoylformate, yielding α-ketoglutarate and d-phenylglycine (d-Phg). Detailed kinetic analysis revealed a range of amine donor and acceptor substrates that allowed the synthesis of enantiopure aromatic d-amino acids at a preparative scale. We also determined the first X-ray crystal structure of d-PhgAT with its bound pyridoxal 5′-phosphate (PLP) cofactor at 2.25 Å resolution. A combination of structural analysis and site-directed mutagenesis of this class III aminotransferase revealed key residues that are potentially involved in the dual substrate recognition, as well as controlling the stereo-inverting behaviour of d-PhgAT. Two arginine residues (Arg34 and Arg407) are involved in substrate recognition within P and O binding pockets respectively. These studies lay the foundation for further enzyme engineering and promote d-PhgAT as a useful biocatalyst for the sustainable production of high value, aromatic d-amino acids. This journal is
Author(s): Serpico A, De Cesare S, Marles-Wright J, Akhtar MK, Loake GJ, Campopiano DJ
Publication type: Article
Publication status: Published
Journal: Catalysis Science and Technology
Year: 2020
Volume: 10
Issue: 19
Pages: 6533-6543
Online publication date: 28/07/2020
Acceptance date: 27/07/2020
Date deposited: 11/12/2020
ISSN (print): 2044-4753
ISSN (electronic): 2044-4761
Publisher: Royal Society of Chemistry
URL: https://doi.org/10.1039/D0CY01391A
DOI: 10.1039/d0cy01391a
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