4-Chlorocatechol 1,2-dioxygenase from the chlorophenol-utilizing Gram-positive Rhodococcus opacus 1CP: crystallization and preliminary crystallographic analysis.
4-Chlorocatechol 1,2-dioxygenase from the chlorophenol-utilizing Gram-positive Rhodococcus opacus 1CP: crystallization and preliminary crystallographic analysis.
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来自利用氯酚的革兰氏阳性红球菌 1CP 的 4-氯儿茶酚 1,2-双加氧酶：结晶和初步晶体学分析。
DOI：
10.1107/s0907444902007242
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
L. Golovleva
中科院分区：
文献类型：
--
作者：
M. Ferraroni;Maria Yolanda Ruiz Tarifa;F. Briganti;A. Scozzafava;S. Mangani;I. Solyanikova;M. Kolomytseva;L. Golovleva
4-氯儿茶酚1,2-双加氧酶（4-ClC1,2DO）来自革兰氏阳性细菌不透明红球菌1CP，一种参与氯芳香族化合物有氧生物降解的酶，已经结晶。4-ClC1,2DO，专门催化4-取代儿茶酚的醇内裂解，而4-取代儿茶酚是各种芳香污染物降解的中间产物，最近被纯化为相应的马来酰乙酸酯。该酶是由α(2)型四元结构的两个相同亚基组成的同型二聚体；每个单体的分子量约为29 kDa，每个同型二聚体含有一个Fe（III）和一个Mn（II）离子。在1.6 M硫酸铵，0.1 M氯化钠，100 mM Tris-HCl pH 9.0, 5-15%甘油中生长的六方晶体在液氮下成功冷冻，在母液溶液中加入30%甘油作为冷冻保护剂。采用EMBL波束线BW7A，在德国汉堡DESY多丽丝存储环上以2.8 A分辨率采集了完整的数据集，采用MAR CCD探测器，波长为1.01 A。晶体属于原始六方空间群P6(3)22，晶胞参数a = 90.4, c = 307.5 a。这是第一个醇内双加氧酶，专门催化革兰氏阳性细菌中氯儿茶酚的裂解，以获得衍射质量的晶体。
4-Chlorocatechol 1,2-dioxygenase (4-ClC1,2DO) from the Gram-positive bacterium Rhodococcus opacus (erythropolis) 1CP, an enzyme involved in the aerobic biodegradation of chloroaromatic compounds, has been crystallized. 4-ClC1,2DO, which specifically catalyzes the intradiol cleavage of 4-substituted catechols, which are intermediates in the degradation of a variety of aromatic pollutants, to the corresponding maleylacetates, has recently been purified to homogeneity. The enzyme is an homodimer composed of two identical subunits in an alpha(2)-type quaternary structure; it has a molecular weight of about 29 kDa per monomer and contains one Fe(III) and one Mn(II) ion per homodimer. Hexagonal crystals grown in 1.6 M ammonium sulfate, 0.1 M sodium chloride, 100 mM Tris-HCl pH 9.0, 5-15% glycerol were successfully frozen under liquid nitrogen, adding 30% glycerol to the mother-liquor solution as a cryoprotectant. A complete data set was collected at 2.8 A resolution using the EMBL beamline BW7A at the DORIS storage ring, DESY, Hamburg, Germany with a MAR CCD detector and a wavelength of 1.01 A. The crystals belong to the primitive hexagonal space group P6(3)22, with unit-cell parameters a = 90.4, c = 307.5 A. This is the first intradiol dioxygenase which specifically catalyzes the cleavage of chlorocatechols in Gram-positive bacteria to give diffraction-quality crystals.
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