恶臭假单胞菌G7中参与萘降解的水杨醛脱氢酶NahF的表达、纯化及初步晶体学研究
Expression, purification and preliminary crystallographic studies of NahF, a salicylaldehyde dehydrogenase from Pseudomonas putida G7 involved in naphthalene degradation.
作者信息
Coitinho Juliana Barbosa, Costa Débora Maria Abrantes, Guimarães Samuel Leite, de Góes Alfredo Miranda, Nagem Ronaldo Alves Pinto
机构信息
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, 31270-901 Belo Horizonte-MG, Brazil.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Jan 1;68(Pt 1):93-7. doi: 10.1107/S174430911105038X. Epub 2011 Dec 24.
Abstract
Pseudomonas putida G7 is one of the most studied naphthalene-degrading species. The nah operon in P. putida, which is present on the 83 kb metabolic plasmid NAH7, codes for enzymes involved in the conversion of naphthalene to salicylate. The enzyme NahF (salicylaldehyde dehydrogenase) catalyzes the last reaction in this pathway. The nahF gene was subcloned into the pET28a(TEV) vector and the recombinant protein was overexpressed in Escherichia coli Arctic Express at 285 K. The soluble protein was purified by affinity chromatography followed by gel filtration. Crystals of recombinant NahF (6×His-NahF) were obtained at 291 K and diffracted to 2.42 Å resolution. They belonged to the hexagonal space group P6(4)22, with unit-cell parameters a = b = 169.47, c = 157.94 Å. The asymmetric unit contained a monomer and a crystallographic twofold axis generated the dimeric biological unit.
摘要
恶臭假单胞菌G7是研究最多的萘降解菌之一。恶臭假单胞菌中的nah操纵子存在于83 kb的代谢质粒NAH7上，编码参与萘转化为水杨酸的酶。NahF酶（水杨醛脱氢酶）催化该途径中的最后一步反应。将nahF基因亚克隆到pET28a(TEV)载体中，并在285 K的大肠杆菌北极特快中过量表达重组蛋白。通过亲和层析随后进行凝胶过滤纯化可溶性蛋白。在291 K下获得重组NahF（6×His-NahF）晶体，衍射分辨率为2.42 Å。它们属于六方空间群P6(4)22，晶胞参数a = b = 169.47，c = 157.94 Å。不对称单元包含一个单体，一个晶体学二重轴产生二聚体生物单元。
相似文献
1
Expression, purification and preliminary crystallographic studies of NahF, a salicylaldehyde dehydrogenase from Pseudomonas putida G7 involved in naphthalene degradation.恶臭假单胞菌G7中参与萘降解的水杨醛脱氢酶NahF的表达、纯化及初步晶体学研究
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Jan 1;68(Pt 1):93-7. doi: 10.1107/S174430911105038X. Epub 2011 Dec 24.
2
Physiological role of the novel salicylaldehyde dehydrogenase NahV in mineralization of naphthalene by Pseudomonas putida ND6.新型水杨醛脱氢酶 NahV 在假单胞菌 ND6 萘矿化中的生理作用。
Microbiol Res. 2011 Dec 20;166(8):643-53. doi: 10.1016/j.micres.2011.01.003. Epub 2011 Mar 3.
3
Expression, purification and crystallization of a novel metagenome-derived salicylaldehyde dehydrogenase from Alpine soil.从高山土壤中新型宏基因组来源的水杨醛脱氢酶的表达、纯化和结晶。
Acta Crystallogr F Struct Biol Commun. 2022 Apr 1;78(Pt 4):161-169. doi: 10.1107/S2053230X22002345. Epub 2022 Mar 28.
4
Structural and kinetic characterization of recombinant 2-hydroxymuconate semialdehyde dehydrogenase from Pseudomonas putida G7.恶臭假单胞菌G7重组2-羟基粘康酸半醛脱氢酶的结构与动力学特性
Arch Biochem Biophys. 2015 Aug 1;579:8-17. doi: 10.1016/j.abb.2015.05.006. Epub 2015 May 29.
5
Bacterial metabolism of naphthalene: construction and use of recombinant bacteria to study ring cleavage of 1,2-dihydroxynaphthalene and subsequent reactions.萘的细菌代谢：构建和使用重组细菌研究1,2-二羟基萘的环裂解及后续反应。
J Bacteriol. 1992 Dec;174(23):7542-54. doi: 10.1128/jb.174.23.7542-7554.1992.
6
Structural and Kinetic Properties of the Aldehyde Dehydrogenase NahF, a Broad Substrate Specificity Enzyme for Aldehyde Oxidation.醛脱氢酶NahF的结构与动力学特性，一种具有广泛底物特异性的醛氧化酶
Biochemistry. 2016 Sep 27;55(38):5453-63. doi: 10.1021/acs.biochem.6b00614. Epub 2016 Sep 13.
7
A combined approach for enhancing the stability of recombinant cis-dihydrodiol naphthalene dehydrogenase from Pseudomonas putida G7 allowed for the structural and kinetic characterization of the enzyme.一种增强恶臭假单胞菌G7重组顺式二氢二醇萘脱氢酶稳定性的联合方法，使得该酶的结构和动力学特性得以表征。
Protein Expr Purif. 2017 Apr;132:50-59. doi: 10.1016/j.pep.2017.01.005. Epub 2017 Jan 9.
8
Identification of nah-1 genes of the Pseudomonas putida naphthalene-degrading NPL-41 plasmid operon.恶臭假单胞菌萘降解NPL-41质粒操纵子nah-1基因的鉴定。
Mol Gen Mikrobiol Virusol. 1999(4):33-6.
9
Effect of temperature, pH, and initial cell number on luxCDABE and nah gene expression during naphthalene and salicylate catabolism in the bioreporter organism Pseudomonas putida RB1353.温度、pH值和初始细胞数量对生物报告菌恶臭假单胞菌RB1353中萘和水杨酸盐分解代谢过程中luxCDABE和nah基因表达的影响。
Appl Environ Microbiol. 2003 Apr;69(4):2209-16. doi: 10.1128/AEM.69.4.2209-2216.2003.
10
Metabolism of dibenzothiophene and naphthalene in Pseudomonas strains: complete DNA sequence of an upper naphthalene catabolic pathway.假单胞菌菌株中二苯并噻吩和萘的代谢：萘分解代谢上游途径的完整DNA序列
J Bacteriol. 1993 Nov;175(21):6890-901. doi: 10.1128/jb.175.21.6890-6901.1993.
引用本文的文献
1
A Novel Acetaldehyde Dehydrogenase with Salicylaldehyde Dehydrogenase Activity from Rhodococcus ruber Strain OA1.来自红球菌属菌株OA1的具有水杨醛脱氢酶活性的新型乙醛脱氢酶。
Curr Microbiol. 2017 Dec;74(12):1404-1410. doi: 10.1007/s00284-017-1333-8. Epub 2017 Aug 28.
2
Evolutionary, computational, and biochemical studies of the salicylaldehyde dehydrogenases in the naphthalene degradation pathway.萘降解途径中水杨醛脱氢酶的进化、计算和生化研究。
Sci Rep. 2017 Feb 24;7:43489. doi: 10.1038/srep43489.
3
iTRAQ-based quantitative proteomic analysis of the global response to 17β-estradiol in estrogen-degradation strain Pseudomonas putida SJTE-1.基于 iTRAQ 的雌激素降解菌恶臭假单胞菌 SJTE-1 对 17β-雌二醇的全局响应的定量蛋白质组学分析。
Sci Rep. 2017 Feb 3;7:41682. doi: 10.1038/srep41682.
4
Structural and kinetic characterization of recombinant 2-hydroxymuconate semialdehyde dehydrogenase from Pseudomonas putida G7.恶臭假单胞菌G7重组2-羟基粘康酸半醛脱氢酶的结构与动力学特性
Arch Biochem Biophys. 2015 Aug 1;579:8-17. doi: 10.1016/j.abb.2015.05.006. Epub 2015 May 29.
5
Improving serodiagnosis of human and canine leishmaniasis with recombinant Leishmania braziliensis cathepsin l-like protein and a synthetic peptide containing its linear B-cell epitope.利用重组巴西利什曼原虫组织蛋白酶L样蛋白和含其线性B细胞表位的合成肽改进人和犬利什曼病的血清学诊断
PLoS Negl Trop Dis. 2015 Jan 8;9(1):e3426. doi: 10.1371/journal.pntd.0003426. eCollection 2015 Jan.
6
Mapping B-cell epitopes for the peroxidoxin of Leishmania (Viannia) braziliensis and its potential for the clinical diagnosis of tegumentary and visceral leishmaniasis.巴西利什曼原虫（维扬尼亚种）过氧化物酶的B细胞表位定位及其在皮肤利什曼病和内脏利什曼病临床诊断中的潜力
PLoS One. 2014 Jun 12;9(6):e99216. doi: 10.1371/journal.pone.0099216. eCollection 2014.
7
Epitope mapping of the HSP83.1 protein of Leishmania braziliensis discloses novel targets for immunodiagnosis of tegumentary and visceral clinical forms of leishmaniasis.巴西利什曼原虫HSP83.1蛋白的表位作图揭示了用于皮肤型和内脏型利什曼病免疫诊断的新靶点。
Clin Vaccine Immunol. 2014 Jul;21(7):949-59. doi: 10.1128/CVI.00151-14. Epub 2014 May 7.
本文引用的文献
1
REFMAC5 for the refinement of macromolecular crystal structures.用于大分子晶体结构精修的REFMAC5
Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67. doi: 10.1107/S0907444911001314. Epub 2011 Mar 18.
2
iMOSFLM: a new graphical interface for diffraction-image processing with MOSFLM.iMOSFLM：一种用于MOSFLM衍射图像处理的新图形界面。
Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):271-81. doi: 10.1107/S0907444910048675. Epub 2011 Mar 18.
3
Features and development of Coot.Coot的特点与发展
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501. doi: 10.1107/S0907444910007493. Epub 2010 Mar 24.
4
Depurinating naphthalene-DNA adducts in mouse skin related to cancer initiation.与癌症起始相关的鼠皮中去嘌呤萘-DNA 加合物。
Free Radic Biol Med. 2009 Oct 1;47(7):1075-81. doi: 10.1016/j.freeradbiomed.2009.07.020. Epub 2009 Jul 18.
5
Phaser crystallographic software.相位结晶学软件。
J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674. doi: 10.1107/S0021889807021206. Epub 2007 Jul 13.
6
Biodegradation aspects of polycyclic aromatic hydrocarbons (PAHs): a review.多环芳烃（PAHs）的生物降解研究综述
J Hazard Mater. 2009 Sep 30;169(1-3):1-15. doi: 10.1016/j.jhazmat.2009.03.137. Epub 2009 Apr 7.
7
Polycyclic aromatic hydrocarbons (PAHs) biodegradation by basidiomycetes fungi, Pseudomonas isolate, and their cocultures: comparative in vivo and in silico approach.担子菌纲真菌、假单胞菌分离株及其共培养物对多环芳烃（PAHs）的生物降解：体内和计算机模拟比较方法
Appl Biochem Biotechnol. 2008 Dec;151(2-3):132-42. doi: 10.1007/s12010-008-8160-0. Epub 2008 Feb 26.
8
Microbial biodegradation of polyaromatic hydrocarbons.多环芳烃的微生物降解
FEMS Microbiol Rev. 2008 Nov;32(6):927-55. doi: 10.1111/j.1574-6976.2008.00127.x. Epub 2008 Jul 24.
9
Physical, kinetic and spectrophotometric studies of a NAD(P)-dependent benzaldehyde dehydrogenase from Pseudomonas putida ATCC 12633.
Biochim Biophys Acta. 2008 Sep;1784(9):1248-55. doi: 10.1016/j.bbapap.2008.04.015. Epub 2008 May 2.
10
DNA adducts formation and induction of apoptosis in rat liver epithelial 'stem-like' cells exposed to carcinogenic polycyclic aromatic hydrocarbons.暴露于致癌多环芳烃的大鼠肝上皮“干细胞样”细胞中DNA加合物的形成及细胞凋亡的诱导
Mutat Res. 2008 Feb 1;638(1-2):122-32. doi: 10.1016/j.mrfmmm.2007.09.004. Epub 2007 Sep 16.
Zotero 插件