Expression, purification, crystallization and preliminary X-ray analysis of para-nitrophenol 4-monooxygenase from Pseudomonas putida DLL-E4
Expression, purification, crystallization and preliminary X-ray analysis of para-nitrophenol 4-monooxygenase from Pseudomonas putida DLL-E4
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恶臭假单胞菌 DLL-E4 对硝基苯酚 4-单加氧酶的表达、纯化、结晶和初步 X 射线分析
DOI：
10.1107/s1744309109032370
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发表时间：
2009-10-01
影响因子：
0.9
通讯作者：
Cui, Zhongli
中科院分区：
文献类型：
--
作者：
Liu, Weidong;Shen, Wenjing;Cui, Zhongli
对硝基苯酚4-单加氧酶(PnpA)在细菌降解对硝基苯酚的过程中起着重要作用，它通过将硝基从芳环上氧化释放出来形成对苯二酚。为了了解该酶功能的结构基础，我们克隆了PnpA，并在大肠杆菌中进行了表达和纯化。以聚乙二醇4000为沉淀剂，采用悬滴气相扩散法制备了PnpA。PnpA晶体属于P2(1)2(1)2(1)空间群，晶胞参数a=54.47，b=77.56，c=209.17埃，衍射分辨率为2.24埃。
Para-nitrophenol 4-monooxygenase (PnpA) plays an important role in bacterial degradation of para-nitrophenol by oxidative release of the nitro group from the aromatic ring to form p-benzoquinone. In order to understand the structural basis of the function of this enzyme, PnpA was cloned, expressed in Escherichia coli and purified. PnpA was crystallized by the hanging-drop vapour-diffusion technique with PEG 4000 as precipitant. The PnpA crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 54.47, b = 77.56, c = 209.17 angstrom, and diffracted to 2.24 angstrom resolution.