Toggle Main Menu Toggle Search

Open Access padlockePrints

The Newcastle University research output collection, currently available on ePrints, will shortly be moving to a new open repository platform, Figshare. To prepare for the data migration we have paused adding new content to ePrints, and will resume once the new repository is launched. During this time you will continue to have access to ePrints (but no new content will appear). We will share updates here when available.

D-Phenylglycine aminotransferase (d-PhgAT)-substrate scope and structural insights of a stereo-inverting biocatalyst used in the preparation of aromatic amino acids

Lookup NU author(s): Dr Jon Marles-WrightORCiD

Downloads

Licence

This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

© 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

Publication metadata

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

Altmetrics

Altmetrics provided by Altmetric

Share