8I99

N-carbamoyl-D-amino-acid hydrolase mutant - M4Th3

Summary for 8I99

• Entry DOI: 10.2210/pdb8i99/pdb
• Descriptor: N-carbamoyl-D-amino-acid hydrolase (1 entity in total)
• Functional Keywords: carbon-nitrogen hydrolase, tetramer, hydrolase
• Biological source: Nitratireductor indicus C115
• Total number of polymer chains: 4
• Total formula weight: 138348.53

Authors

Hu, J.M.,Ni, Y.,Xu, G.C. (deposition date: 2023-02-06, release date: 2023-03-22, Last modification date: 2024-05-29)

Primary citation

Hu, J.,Chen, X.,Zhang, L.,Zhou, J.,Xu, G.,Ni, Y.
Engineering the Thermostability of a d-Carbamoylase Based on Ancestral Sequence Reconstruction for the Efficient Synthesis of d-Tryptophan.
J.Agric.Food Chem., 71:660-670, 2023

PubMed Abstract: Employing ancestral sequence reconstruction and consensus sequence analysis, the thermostability of a novel d-carbamoylase derived from (HyuC) was engineered through greedy-oriented iterative combinatorial mutagenesis. A mutant S202P/E208D/R277L (M4Th3) was obtained with significantly elevated thermostability. M4Th3 has a half-life of 36.5 h at 40 °C, about 28.5 times of 1.3 h of its parent M4. For the reaction at 40 °C, M4Th3 can catalyze 10 mM -carbamoyl-d-tryptophan to produce d-tryptophan with a conversion ratio of 96.4% after 12 h, which is significantly higher than 64.1% of M4. MD simulation reveals that new hydrogen bonds emerging from E208D on the surface can increase the hydrophobicity of the protein, leading to improved stability. More importantly, R277L could contribute to enhanced interface stability of homodimeric M4. This study provides a thermostable d-carbamoylase for the "hydantoinase process", which has potential in the industrial synthesis of optically pure natural and non-natural amino acids.

PubMed: 36541894
DOI: 10.1021/acs.jafc.2c07781

Experimental method

X-RAY DIFFRACTION (3.17 Å)