8c46

Publication Abstract from PubMed

N-carbamoyl-beta-alanine amidohydrolase (CbetaAA) 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 CbetaAA-encoding gene from Rhizobium radiobacter strain MDC 8606 was cloned and overexpressed in Escherichia coli. The purified recombinant enzyme (RrCbetaAA) showed a specific activity of 14 U/mg using N-carbamoyl-beta-alanine as a substrate with an optimum activity at 55 degrees C and pH 8.0. In this work, we report also the first prokaryotic CbetaAA structure at a resolution of 2.0 A. A discontinuous catalytic domain and a dimerization 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 RrCbetaAA from R. radiobacter MDC 8606 for the industrial production of L-alpha-, L-beta- and L-gamma-amino acids. The structural analysis provides new insights on enzyme-substrate interaction, which shed light on engineering of CbetaAAs for high catalytic activity and broad substrate specificity.

Structural and biochemical characterisation of the N-carbamoyl-beta-alanine amidohydrolase from Rhizobium radiobacter MDC 8606.,Paloyan A, Sargsyan A, Karapetyan MD, Hambardzumyan A, Kocharov S, Panosyan H, Dyukova K, Kinosyan M, Kreuger A, Piergentili C, Stanley WA, Djoko KY, Basle A, Marles-Wright J, Antranikian G FEBS J. 2023 Aug 27. doi: 10.1111/febs.16943. PMID:37634202^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.