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Biochemistry of bacterial dissimilatory metal reduction

细菌异化金属还原的生物化学

基本信息

批准号：
131646117
负责人：
Professor Dr. Johannes Gescher
金额：
--
依托单位：
Institut für Technische Mikrobiologie V-7
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2009
资助国家：
德国
起止时间：
2008-12-31 至 2015-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/131646117?language=en
关键词：
Biochemistry bacterial dissimilatory metal reduction

项目摘要

Dissimilatory metal reduction is a respiratory process that operates in various Bacteria and Archaea. These microorganisms are able to use oxidized metals like Fe(III), Mn(IV), Cr(VI), or U(VI) as terminal electron acceptors of their anaerobic electron transport chains. Shewanella oneidensis is one of the two model organisms to study this process. However, even after decades of research on dissimilatory metal reduction, the essential minimal protein set for the electron transport to metallic electron acceptors is still unknown. I am pursuing a novel synthetic biology based approach to tackle this problem. Therefore, the goal of the proposed study is to convert Escherichia coli into a dissimilatory metal reducer through expression of potential key genes from S. oneidensis. Using this technique we have already established a strain that is able to respire with chelated Fe(III) as terminal electron acceptor and have thereby shown the applicability of the method. Via the insertion of further genes for possible compounds of the electron transport chain from S. oneidensis we want to proceed to the next level, being to establish an electron transfer to crystalline iron and manganese forms. We will detect potential key players for the reduction chain to ferric iron and manganese using several parallel approaches, which include mass spectrometry, purification and following characterization as well as targeted gene deletions. A series of proof of principle experiments for the reduction chain to ferric iron were already conducted. These in vitro experiments revealed interprotein electron transfer reactions and were also verified by in vivo experiments. The designed E. coli strains are supposed to be further used to address questions of protein localization and complex formation and as a platform for heterologues expression of targets for further crystallization studies.

异化金属还原是一种在各种细菌和古生菌中运行的呼吸过程。这些微生物能够利用铁(III)、锰(IV)、铬(VI)或铀(VI)等氧化金属作为其厌氧电子传递链的末端电子受体。单氏希瓦氏菌是研究这一过程的两种模式生物之一。然而，即使经过几十年的异化金属还原研究，电子传递到金属电子受体所必需的最小蛋白质集仍然是未知的。我正在寻求一种基于合成生物学的新方法来解决这个问题。因此，本研究的目的是通过表达金黄色葡萄球菌潜在的关键基因，将大肠杆菌转化为异化的金属还原体。利用这一技术，我们已经建立了一株能够以螯合Fe(III)作为末端电子受体进行呼吸的菌株，从而证明了该方法的适用性。通过插入更多可能的电子传输链化合物的基因，我们希望进入下一个水平，即建立一个电子转移到结晶铁和锰的形式。我们将使用几种平行的方法来发现铁铁和锰还原链的潜在关键角色，这些方法包括质谱学、纯化和后续表征以及靶向基因缺失。对铁还原链进行了一系列的原理性实验论证。这些体外实验揭示了蛋白质间的电子转移反应，也得到了体内实验的验证。设计的大肠杆菌菌株有望进一步用于解决蛋白质定位和复合体形成的问题，并作为进一步结晶研究的靶标异源表达的平台。