来自波氏潘多拉菌B-356的顺式联苯-2,3-二氢二醇-2,3-脱氢酶的表达、纯化、结晶及初步晶体学研究
Expression, purification, crystallization and preliminary crystallographic studies of cis-biphenyl-2,3-dihydrodiol-2,3-dehydrogenase from Pandoraea pnomenusa B-356.
作者信息
Patil Dipak N, Tomar Shailly, Sylvestre Michel, Kumar Pravindra
机构信息
Department of Biotechnology, Indian Institute of Technology, Roorkee, Roorkee 247 667, India.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Nov 1;66(Pt 11):1517-20. doi: 10.1107/S1744309110036894. Epub 2010 Oct 29.
Abstract
cis-Biphenyl-2,3-dihydrodiol-2,3-dehydrogenase (BphB) is involved in the aerobic biodegradation of biphenyl and polychlorinated biphenyls. BphB from Pandoraea pnomenusa strain B-356 was overexpressed in Escherichia coli, purified to homogeneity and crystallized. Crystals were obtained by the sitting-drop vapour-diffusion method using polyethylene glycol 3350 and 0.2 M sodium malonate. A BphB crystal diffracted to 2.8 Å resolution and belonged to space group P4(3)2(1)2, with unit-cell parameters a = b = 75.2, c = 180.4 Å. Preliminary crystallographic analysis indicated the presence of two molecules in the asymmetric unit, giving a Matthews coefficient of 2.2 Å(3) Da(-1) and a solvent content of 44%.
摘要
顺式联苯-2,3-二氢二醇-2,3-脱氢酶（BphB）参与联苯和多氯联苯的好氧生物降解。来自潘多拉菌属pnomenusa菌株B-356的BphB在大肠杆菌中过表达，纯化至同质并结晶。通过坐滴气相扩散法，使用聚乙二醇3350和0.2 M丙二酸钠获得晶体。一个BphB晶体衍射至2.8 Å分辨率，属于空间群P4(3)2(1)2，晶胞参数a = b = 75.2，c = 180.4 Å。初步晶体学分析表明不对称单元中存在两个分子，马修斯系数为2.2 Å(3) Da(-1)，溶剂含量为44%。
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