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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >4-Chlorocatechol 1,2-dioxygenase from the chlorophenol-utilizing Gram-positive Rhodococcus opacus 1CP: crystallization and preliminary crystallographic analysis.
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4-Chlorocatechol 1,2-dioxygenase from the chlorophenol-utilizing Gram-positive Rhodococcus opacus 1CP: crystallization and preliminary crystallographic analysis.

机译：4-Chlorocatechol 2-dioxygenase从chlorophenol-utilizing革兰氏阳性红球菌属蔽光的1 cp:结晶和初步的晶体分析。

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摘要

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.
机译：4-Chlorocatechol 2-dioxygenase (4-ClC1 2)革兰氏阳性细菌红球菌属蔽光的(erythropolis) 1 cp,一种酶参与好氧生物降解的chloroaromatic化合物,结晶。特别是催化intradiol劈理的4-substituted儿茶酚,中间体的降解多种芳香族污染物,相应的maleylacetates,最近被净化同质性。两个相同的亚基的α(2)类型四级结构;约29 kDa /单体和包含一个铁(III)和一个锰(II)每为离子。六角晶体生长在1.6铵硫酸,0.1 M氯化钠,100毫米Tris-HClpH值9.0,5 - 15%甘油被成功冻结在液态氮,添加30%甘油母液的解决方案作为冷冻保护剂。2.8收集完整的数据集解决使用EMBL beamline BW7A在多丽丝存储戒指,谜底,德国汉堡3月CCD探测器和一个波长的1.01。晶体属于原始的六角形空间集团P6(3) 22日,晶胞参数=90.4, c = 307.5。加双氧酶,特别是催化乳沟chlorocatechols革兰氏阳性细菌给diffraction-quality晶体。

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