天津科技大学学报2017，Vol.32Issue(2)：19-23,29,6.DOI:10.13364/j.issn.1672-6510.20160024
睾丸酮丛毛单胞菌CNB1中2-氨基-5-氯粘康酸半醛脱氢酶的表达纯化与结晶
Expression,Purification and Crystallization of 2-amino-5-chloromuconic semialdehyde dehydrogenase from Comamonas testosteroniCNB1
摘要
Abstract
Although nitrobenzene and its derivatives(NBDs)are important industrial materials,they can cause severe dam-age to the environment.Physical,chemical and biological methods are currently used to degrade residual NBDs in the envi-ronment,among which the biological method is considered to be the most promising,given its advantages such as low cost, little secondary pollution,and ecological restoration. 4-chloronitrobenzene(4,CNB),one of the highly toxic and dangerous NBDs,can cause persistent physiological damage to human beings via direct or indirect contact.The 2-amino-5-chloromuconic semialdehyde dehydrogenase(CnbD)from Comamonas testosteroni CNB1 is an important component in-volved in 4,CNB degradation.The crystal structure of CnbD can help us to understand the mechanism of this enzyme and make full use of CnbD to degrade 4,CNB in the environment.ThecnbD gene was cloned into the prokaryotic expression vector pET-28,a(+)and the CnbD protein was overexpressed in Escherichia coli.The CnbD protein was purified with Ni2+-chelating affinity chromatography and gel filtration chromatography.Protein crystals were cultivated at 289,K and diffracted to resolution of 0.18,nm and 0.22,nm respectively at the wavelength of 0.10,nm in SSRF.With the molecular-replacement (MR)method,the three-dimensional structure of the protein was successfully determined.关键词
2-氨基-5-氯粘康酸半醛脱氢酶/对氯硝基苯/睾丸酮丛毛单胞菌/蛋白晶体/分子置换法Key words
2-amino-5-chloromuconic semialdehyde dehydrogenase/4-chloronitrobenzene/Comamonas testosteroni/protein crystals/molecular-replacement分类
生物科学引用本文复制引用
陈文成,谢元美,刘祥,赵树欣..睾丸酮丛毛单胞菌CNB1中2-氨基-5-氯粘康酸半醛脱氢酶的表达纯化与结晶[J].天津科技大学学报,2017,32(2):19-23,29,6.基金项目
国家自然科学基金资助项目(31200641) （31200641）
