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亚铁还原黏细菌 Anaeromyxobacter dehalogenans 2CP-C 利用延胡索酸盐和柠檬酸铁生长时的综合蛋白质组分析

Comprehensive proteome profiling of the Fe(III)-reducing myxobacterium Anaeromyxobacter dehalogenans 2CP-C during growth with fumarate and ferric citrate.

机构信息

Fakultät für Physik, Universität Bielefeld, Bielefeld, Germany.

出版信息

Proteomics. 2010 Apr;10(8):1673-84. doi: 10.1002/pmic.200900687.

Abstract

Anaeromyxobacter dehalogenans is a microaerophilic member of the delta-proteobacteria which is able to utilize a wide range of electron acceptors, including halogenated phenols, U(VI), Fe(III), nitrate, nitrite, oxygen and fumarate. To date, the knowledge regarding general metabolic activities of this ecologically relevant bacterium is limited. Here, we present a first systematic 2-D reference map of the soluble A. dehalogenans proteome in order to provide a sound basis for further proteomic studies as well as to gain first global insights into the metabolic activities of this bacterium. Using a combination of 2-DE and MALDI-TOF-MS, a total of 720 proteins spots were identified, representing 559 unique protein species. Using the proteome data, altogether 50 metabolic pathways were found to be expressed during growth with fumarate as primary electron acceptor. An analysis of the pathways revealed an extensive display of enzymes involved in the catabolism and anabolism of a variety of amino acids, including the unexpected fermentation of lysine to butyrate. Moreover, using the reference gel as basis, a semi-quantitative analysis of protein expression changes of A. dehalogenans during growth with ferric citrate as electron acceptor was conducted. The adaptation to Fe(III) reducing conditions involved the expression changes of a total of 239 proteins. The results suggest that the adaptation to Fe(III) reductive conditions involves an increase in metabolic flux through the tricarboxylic acid cycle, which is fueled by an increased catabolism of amino acids.

摘要

厌氧食酸菌是δ变形菌纲的一种微好氧菌,能够利用多种电子受体,包括卤代酚、U(VI)、Fe(III)、硝酸盐、亚硝酸盐、氧气和延胡索酸。迄今为止,人们对这种具有生态相关性的细菌的一般代谢活性的了解有限。在这里,我们首次呈现了一个可溶厌氧食酸菌蛋白质组的 2-D 参考图谱,以便为进一步的蛋白质组学研究提供坚实的基础,并首次全面了解该细菌的代谢活性。使用 2-DE 和 MALDI-TOF-MS 的组合,共鉴定出 720 个蛋白质点,代表 559 个独特的蛋白质种类。使用蛋白质组数据,总共发现 50 条代谢途径在以延胡索酸为主要电子受体的生长过程中表达。对途径的分析表明,参与各种氨基酸的分解代谢和合成代谢的酶广泛存在,包括赖氨酸意外发酵为丁酸盐。此外,使用参考凝胶作为基础,对以柠檬酸铁作为电子受体时厌氧食酸菌的蛋白质表达变化进行了半定量分析。适应 Fe(III)还原条件涉及总共 239 种蛋白质的表达变化。结果表明,适应 Fe(III)还原条件涉及通过三羧酸循环增加代谢通量,这是由氨基酸分解代谢增加提供动力的。

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