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 P4₁2₁2, 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.

中文翻译：

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来自伯克霍尔德氏菌的 N-乙酰基-(R)-β-苯丙氨酸酰基转移酶的表达、纯化和结晶。AJ110349 和 Variovorax sp。AJ110348 和伯克霍尔德氏菌酶的结构测定

N-乙酰基-( R )-β-苯丙氨酸酰基转移酶是水解N-乙酰基-( R )-β-苯丙氨酸的酰胺键生成对映体纯的( R )-β-苯丙氨酸的酶。在以前的研究中，Burkholderia sp。AJ110349 和Variovorax sp。AJ110348 被分离为 ( R )-对映异构体特异性N-乙酰基-( R )-β-苯丙氨酸酰基转移酶产生生物和来自伯克霍尔德氏菌的天然酶的特性sp。AJ110349 进行了表征。在这项研究中，进行了结构分析，以研究源自这两种生物的酶的结构-功能关系。重组N-乙酰基-( R )-β-苯丙氨酸酰基转移酶采用悬滴气相扩散法在多重结晶溶液条件下结晶。伯克霍尔德氏菌酶的晶体属于空间群P 4 ₁ 2 ₁ 2，晶胞参数a = b = 112.70–112.97，c= 341.50–343.32 Å，并且可能在不对称单元中包含两个亚基。通过Se-SAD方法解析晶体结构，表明不对称单元中的两个亚基形成二聚体。每个亚基由三个结构域组成，它们与来自Paracoccus sp.的N , N-二甲基甲酰胺酶的大亚基的相应结构域显示出结构相似性。应变DMF。Variovorax酶的晶体生长为孪晶，不适用于结构测定。使用尺寸排阻色谱法和在线静态光散射分析，N-乙酰基-( R )-β-苯丙氨酸酰基转移酶在溶液中被澄清为二聚体。