Regulation of substrate dependent expression and activity from dehalogenating enzymes in Dehalococcoides mccartyi strain CBDB1

Dehalococcoides mccartyi 菌株 CBDB1 中脱卤酶的底物依赖性表达和活性的调节

• 批准号: 191845007
• 负责人: Professor Dr. Martin von Bergen
• 金额: --
• 依托单位: Department Molekulare Systembiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/191845007?language=en
• 关键词: Regulation; substrate; dependent; expression; activity

The biochemistry of organohalide respiration is still elusive because some of the species capable of it do not provide sufficient biomass for classical biochemistry on purified enzymes. The improvement of sensitivity in mass spectrometry of peptides together with available genomic information will enhance the study of the biochemical principles of organohalide respirers. The species Sulfurospirillum multivorans, Dehalococcoides sp. CBDB1, DCMB5 and BTF08 will be provided by the subproject partners 1, 2, 3 and 4 within the framework of this Research Unit and cultivated with different electron donors, electron acceptors and carbon sources. The limited biomass of the Dehalococcoides spp. requires optimisation of both protein extraction protocols and measurement by high-resolution mass spectrometers. Shotgun proteomic analyses will provide a deeper insight into the proteins involved and thus the corresponding biochemical pathways. Among the different organohalide respiring species a wide variety of reductive dehalogenase genes are found and neither their substrate specificity nor the regulation of their synthesis in response to the substrate is understood. An accurate detection and quantification of the different gene products of the reductive dehalogenases will be performed by selected reaction monitoring – mass spectrometry (srm-MS). This targeted proteomics approach uses standard peptides labelled with heavy isotopes for quantification of peptides of interest in digested protein extracts by qtrap-MS. The specific quantification will yield insights into the correlation between substrate, culture conditions and the biochemical pathways involved as well as the expression of orthologous reductive dehalogenases.

有机卤化物呼吸的生物化学仍然是难以捉摸的，因为一些能够呼吸的物种不能提供足够的生物量用于纯化酶的经典生物化学。肽质谱灵敏度的提高以及可用的基因组信息将促进有机卤化物呼吸器的生化原理的研究。Sulfurospirillum multivorans、Dehalococcoides sp. CBDB 1、DCMB 5和BTF 08物种将由本研究单元框架内的子项目合作伙伴1、2、3和4提供，并使用不同的电子供体、电子受体和碳源进行培养。Dehalococcoides spp.的有限生物量。需要优化蛋白质提取方案和高分辨率质谱仪的测量。鸟枪蛋白质组学分析将提供更深入的了解所涉及的蛋白质，从而相应的生化途径。在不同的有机卤化物呼吸物种中，发现了各种各样的还原脱卤酶基因，并且既不了解它们的底物特异性，也不了解它们响应于底物的合成的调节。还原性脱卤酶的不同基因产物的准确检测和定量将通过选择反应监测-质谱法（srm-MS）进行。这种靶向蛋白质组学方法使用重同位素标记的标准肽，通过qtrap-MS对消化的蛋白质提取物中的感兴趣肽进行定量。具体的定量将深入了解底物、培养条件和所涉及的生化途径之间的相关性，以及邻苯二甲酸还原脱卤酶的表达。

项目成果

A Retentive Memory of Tetrachloroethene Respiration in Sulfurospirillum halorespirans - involved Proteins and a possible link to Acetylation of a Two-Component Regulatory System.

• DOI: 10.1016/j.jprot.2018.03.030
• 发表时间: 2018-06
• 期刊: Journal of proteomics
• 影响因子: 3.3
• 作者: Dominique Türkowsky;Jens Esken;Tobias Goris;T. Schubert;G. Diekert;N. Jehmlich;M. von Bergen

Thermal proteome profiling allows quantitative assessment of interactions between tetrachloroethene reductive dehalogenase and trichloroethene.

• DOI: 10.1016/j.jprot.2018.05.018
• 发表时间: 2019-02
• 期刊: Journal of proteomics
• 影响因子: 3.3
• 作者: Dominique Türkowsky;Patrick Lohmann;Marie Mühlenbrink;T. Schubert;L. Adrian;Tobias Goris;N. Jehmlich;M. von Bergen

Proteomic dataset of the organohalide-respiring bacterium Dehalococcoides mccartyi strain CBDB1 grown on hexachlorobenzene as electron acceptor

以六氯苯为电子受体生长的有机卤化物呼吸细菌 Dehalococcoides mccartyi 菌株 CBDB1 的蛋白质组数据集

• DOI: 10.1016/j.dib.2016.02.037
• 发表时间: 2016
• 期刊: Data in Brief
• 影响因子: 1.2
• 作者: Schiffmann CL;Otto W;Hansen R;Nielsen PH;Adrian L;Seifert J;von Bergen M;Jehmlich N
• 通讯作者: Jehmlich N

Comparison of targeted peptide quantification assays for reductive dehalogenases by selective reaction monitoring (SRM) and precursor reaction monitoring (PRM)

• DOI: 10.1007/s00216-013-7451-7
• 发表时间: 2014-01-01
• 期刊: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
• 影响因子: 4.3
• 作者: Schiffmann, Christian;Hansen, Rasmus;Seifert, Jana
• 通讯作者: Seifert, Jana