恶臭假单胞菌DLL-E4对硝基苯酚4-单加氧酶的表达、纯化、结晶及初步X射线分析
Expression, purification, crystallization and preliminary X-ray analysis of para-nitrophenol 4-monooxygenase from Pseudomonas putida DLL-E4.
作者信息
Liu Weidong, Shen Wenjing, Zhao Xiaoli, Cao Hui, Cui Zhongli
机构信息
Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, 210095 Nanjing, People's Republic of China.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Oct 1;65(Pt 10):1004-6. doi: 10.1107/S1744309109032370. Epub 2009 Sep 23.
Abstract
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 A, and diffracted to 2.24 A resolution.
摘要
对硝基苯酚4-单加氧酶（PnpA）在细菌通过将硝基从芳环上氧化释放以形成对苯醌来降解对硝基苯酚的过程中发挥着重要作用。为了理解该酶功能的结构基础，PnpA被克隆、在大肠杆菌中表达并纯化。采用悬滴气相扩散技术，以聚乙二醇4000作为沉淀剂使PnpA结晶。PnpA晶体属于空间群P2(1)2(1)2(1)，晶胞参数a = 54.47、b = 77.56、c = 209.17 Å，衍射分辨率达到2.24 Å。
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