8I59

N-acetyl-(R)-beta-phenylalanine acylase, selenomethionyl derivative

Summary for 8I59

• Entry DOI: 10.2210/pdb8i59/pdb
• Related: 8HUY
• Descriptor: N-acetyl-(R)-beta-phenylalanine acylase (1 entity in total)
• Functional Keywords: beta-phenylalanine, amidohydrolase, hydrolase
• Biological source: Burkholderia sp.
• Total number of polymer chains: 2
• Total formula weight: 167083.28

Authors

Kato, Y.,Natsume, R. (deposition date: 2023-01-24, release date: 2023-03-15, Last modification date: 2024-10-16)

Primary citation

Kato, Y.,Kawasaki, H.,Nakamatsu, T.,Matsuda, N.,Natsume, R.
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.
Acta Crystallogr.,Sect.F, 79:70-78, 2023

PubMed Abstract: N-Acetyl-(R)-β-phenylalanine acylase is an enzyme that hydrolyzes the amide bond of N-acetyl-(R)-β-phenylalanine to produce enantiopure (R)-β-phenylalanine. In previous studies, Burkholderia sp. AJ110349 and Variovorax sp. AJ110348 were isolated as (R)-enantiomer-specific N-acetyl-(R)-β-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)-β-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 P422, with unit-cell parameters a = b = 112.70-112.97, c = 341.50-343.32 Å, 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)-β-phenylalanine acylases were clarified to be dimeric in solution.

PubMed: 36862095
DOI: 10.1107/S2053230X23000730

Experimental method

X-RAY DIFFRACTION (3.21 Å)