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Structural and biochemical characterisation of the N-carbamoyl-β-alanine amidohydrolase from Rhizobium radiobacter MDC 8606

Lookup NU author(s): Dr Ani Paloyan, Dr Armen Sargsyan, Dr Cecilia Piergentili, Dr Will Stanley, Dr Arnaud Basle, Dr Jon Marles-WrightORCiD



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


© 2023 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.N-carbamoyl-β-alanine amidohydrolase (CβAA) constitutes one of the most important groups of industrially relevant enzymes used in the production of optically pure amino acids and derivatives. In this study, a CβAA-encoding gene from Rhizobium radiobacter strain MDC 8606 was cloned and overexpressed in Escherichia coli. The purified recombinant enzyme (RrCβAA) showed a specific activity of 14 U·mg−1 using N-carbamoyl-β-alanine as a substrate with an optimum activity at 55 °C and pH 8.0. In this work, we report also the first prokaryotic CβAA structure at a resolution of 2.0 Å. A discontinuous catalytic domain and a dimerisation domain attached through a flexible hinge region at the domain interface have been revealed. We identify key ligand binding residues, including a conserved glutamic acid (Glu131), histidine (H385) and arginine (Arg291). Our results allowed us to explain the preference of the enzyme for linear carbamoyl substrates, as large and branched carbamoyl substrates cannot fit in the active site of the enzyme. This work envisages the use of RrCβAA from R. radiobacter MDC 8606 for the industrial production of L-α-, L-β- and L-γ-amino acids. The structural analysis provides new insights on enzyme–substrate interaction, which shed light on engineering of CβAAs for high catalytic activity and broad substrate specificity.

Publication metadata

Author(s): Paloyan A, Sargsyan A, Karapetyan MD, Hambardzumyan A, Kocharov S, Panosyan H, Dyukova K, Kinosyan M, Krueger A, Piergentili C, Stanley WA, Djoko KY, Basle A, Marles-Wright J, Antranikian G

Publication type: Article

Publication status: Published

Journal: FEBS Journal

Year: 2023

Volume: 290

Issue: 23

Pages: 5566-5580

Print publication date: 01/12/2023

Online publication date: 27/08/2023

Acceptance date: 25/08/2023

Date deposited: 20/09/2023

ISSN (print): 1742-464X

ISSN (electronic): 1742-4658

Publisher: John Wiley and Sons Inc


DOI: 10.1111/febs.16943

PubMed id: 37634202


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Funder referenceFunder name
BB/N005570/1Biotechnology and Biological Sciences Research Council (BBSRC)
FAST Travel Grant for Collaborative Research
Newcastle University