8i5a | pdb_00008i5a
N-acetyl-(R)-beta-phenylalanine acylase, 2.75 angstrom resolution
Structural highlights
Publication Abstract from PubMedN-Acetyl-(R)-beta-phenylalanine acylase is an enzyme that hydrolyzes the amide bond of N-acetyl-(R)-beta-phenylalanine to produce enantiopure (R)-beta-phenylalanine. In previous studies, Burkholderia sp. AJ110349 and Variovorax sp. AJ110348 were isolated as (R)-enantiomer-specific N-acetyl-(R)-beta-phenylalanine acylase-producing organisms and the properties of the native enzyme from Burkholderia sp. AJ110349 were characterized. In this study, structural analyses were carried out in order to investigate the structure-function relationships of the enzymes derived from both organisms. The recombinant N-acetyl-(R)-beta-phenylalanine acylases were crystallized by the hanging-drop vapor-diffusion method under multiple crystallization solution conditions. The crystals of the Burkholderia enzyme belonged to space group P4(1)2(1)2, with unit-cell parameters a = b = 112.70-112.97, c = 341.50-343.32 A, and were likely to contain two subunits in the asymmetric unit. The crystal structure was solved by the Se-SAD method, suggesting that two subunits in the asymmetric unit form a dimer. Each subunit was composed of three domains, and they showed structural similarity to the corresponding domains of the large subunit of N,N-dimethylformamidase from Paracoccus sp. strain DMF. The crystals of the Variovorax enzyme grew as twinned crystals and were not suitable for structure determination. Using size-exclusion chromatography with online static light-scattering analysis, the N-acetyl-(R)-beta-phenylalanine acylases were clarified to be dimeric in solution. Expression, purification and crystallization of N-acetyl-(R)-beta-phenylalanine acylases derived from Burkholderia sp. AJ110349 and Variovorax sp. AJ110348 and structure determination of the Burkholderia enzyme.,Kato Y, Kawasaki H, Nakamatsu T, Matsuda N, Natsume R Acta Crystallogr F Struct Biol Commun. 2023 Mar 1;79(Pt 3):70-78. doi: , 10.1107/S2053230X23000730. Epub 2023 Feb 23. PMID:36862095[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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