树脂糖假单胞菌CA10菌株咔唑1,9a-双加氧酶末端加氧酶组分的结晶及初步晶体学分析
Crystallization and preliminary crystallographic analysis of the terminal oxygenase component of carbazole 1,9a-dioxygenase of Pseudomonas resinovorans strain CA10.
作者信息
Nam Jeong Won, Fujimoto Zui, Mizuno Hiroshi, Yamane Hisakazu, Yoshida Takako, Habe Hiroshi, Nojiri Hideaki, Omori Toshio
机构信息
Biotechnology Research Center, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
出版信息
Acta Crystallogr D Biol Crystallogr. 2002 Aug;58(Pt 8):1350-2. doi: 10.1107/s0907444902009150. Epub 2002 Jul 20.
The terminal oxygenase component (CarAa) of carbazole 1,9a-dioxygenase from Pseudomonas resinovorans strain CA10 was crystallized at 293 K using the sitting-drop vapour-diffusion method under the following conditions: 0.1 M sodium citrate pH 5.6 in the presence of 0.5 M ammonium sulfate and 1.0 M lithium sulfate. By using additive reagents with the crystallizing condition, improved diffraction was obtained from the crystals. Preliminary X-ray diffraction analysis indicated that CarAa crystals are hexagonal and belong to space group P6(2) or P6(4), with unit-cell parameters a = b = 244.5, c = 65.7 A, alpha = beta = 90.0, gamma = 120.0 degrees. Diffraction data were collected to 3.0 A resolution. The V(M) value is 2.16 A(3) Da(-1), which indicates a solvent content of 43.0%. This is the first report of crystallization of the terminal oxygenase component of an angular-type dioxygenase.
利用坐滴气相扩散法，在以下条件下于293 K对树脂糖假单胞菌CA10菌株咔唑1,9a -双加氧酶的末端加氧酶组分（CarAa）进行了结晶：在存在0.5 M硫酸铵和1.0 M硫酸锂的情况下，使用0.1 M柠檬酸钠（pH 5.6）。通过在结晶条件中使用添加剂试剂，从晶体获得了更好的衍射效果。初步X射线衍射分析表明，CarAa晶体为六方晶系，属于空间群P6(2)或P6(4)，晶胞参数a = b = 244.5，c = 65.7 Å，α = β = 90.0，γ = 120.0°。衍射数据收集至3.0 Å分辨率。V(M)值为2.16 ų Da⁻¹，这表明溶剂含量为4３.0%。这是对角型双加氧酶末端加氧酶组分结晶的首次报道。
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