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8i99

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

Structural highlights

8i99 is a 4 chain structure with sequence from Nitratireductor indicus C115. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 3.17Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

K2NMS4_9HYPH

Publication Abstract from PubMed

Employing ancestral sequence reconstruction and consensus sequence analysis, the thermostability of a novel d-carbamoylase derived from Nitratireductor indicus (NiHyuC) 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 degrees C, about 28.5 times of 1.3 h of its parent M4. For the reaction at 40 degrees C, M4Th3 can catalyze 10 mM N-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.

Engineering the Thermostability of a d-Carbamoylase Based on Ancestral Sequence Reconstruction for the Efficient Synthesis of d-Tryptophan.,Hu J, Chen X, Zhang L, Zhou J, Xu G, Ni Y J Agric Food Chem. 2023 Jan 11;71(1):660-670. doi: 10.1021/acs.jafc.2c07781. Epub , 2022 Dec 21. PMID:36541894^[1]

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

References

↑ 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. 2023 Jan 11;71(1):660-670. PMID:36541894 doi:10.1021/acs.jafc.2c07781