Synthetic engineering of conductive biofilm development in the y-proteobacterium Shewanella oneidensis

y-变形杆菌 Shewanella oneidensis 中导电生物膜发育的合成工程

• 批准号: 451681210
• 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
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/451681210?language=en
• 关键词: Synthetic; engineering; conductive; biofilm; development

The aim of this proposal is to understand the boundary conditions of conductive biofilm development by Shewanella oneidensis on electrode surfaces and the synthetic development of an advanced ability of the organism to endogenously build these current producing structures. S. oneidensis is the best understood model organism regarding extracellular electron transfer and the ability to thrive with insoluble electron acceptors. Still, the achievable electron transfer rates with these electron acceptors are several fold lower compared to the other model organism for extracellular electron transfer – Geobacter sulfurreducens. Our research builds upon the hypothesis that the reduction of electron acceptors like hematite or the ability to produce a current in a bioelectrochemical system is correlated to the ability to produce conductive biofilms. This is corroborated by the ability of G. sulfurreducens cells to integrate conductive structures in its extracellular polymeric substance. In the proposed work we will fundamentally establish how biofilm production on anode surfaces by S. oneidensis can be steered by the addition of abiotic or biotic conductive structures. To this end we will use a recently establish microfluidic platform that was advanced for use with bioelectrochemical systems. We want to understand how the organism reacts to the integration of this material and what the maximum current densities are that can be established using these exogenous current collectors. This maximum current density will be the benchmark that we want to reach when we engineer S. oneidensis to endogenously produce conductive structures within its extracellular polymeric material. We will export outer membrane cytochromes into the matrix and will synthetically connect them via isopeptide bonds to other proteins on the surface of the cell. To reach this goal we will use the recently established SpyTag/SpyCatcher technology that allows the posttranslational connection of proteins.

该提案的目的是了解电极表面上希瓦氏菌oneidensis的导电生物膜发展的边界条件，以及生物体内源性构建这些电流产生结构的高级能力的合成发展。S. oneidensis是关于细胞外电子转移和利用不溶性电子受体生长的能力的最好理解的模式生物。尽管如此，这些电子受体可实现的电子转移速率比细胞外电子转移的另一种模式生物-Gesterylsulfurreducens低几倍。我们的研究建立在这样的假设之上，即电子受体如赤铁矿的还原或在生物电化学系统中产生电流的能力与产生导电生物膜的能力相关。这是由G.硫还原细胞以在其细胞外聚合物中整合导电结构。在本研究中，我们将从根本上确定S.可以通过添加非生物或生物传导结构来操纵oneidensis。为此，我们将使用最近建立的微流体平台，该平台是先进的，可用于生物电化学系统。我们想了解生物体如何对这种材料的整合做出反应，以及使用这些外源集电器可以建立的最大电流密度是多少。这个最大电流密度将是我们在设计S时希望达到的基准。一种是在其细胞外聚合物材料内内源性地产生导电结构。我们将输出外膜细胞色素到基质中，并通过异肽键将它们合成连接到细胞表面的其他蛋白质上。为了实现这一目标，我们将使用最近建立的SpyTag/SpyCatcher技术，该技术允许蛋白质的翻译后连接。