Direct Stimulation of Bacterial Metabolism with Applied Electrical Current

施加电流直接刺激细菌代谢

• 批准号: 8627970
• 负责人: Drew Carl MacKellar
• 金额: $ 5.33万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8627970
• 关键词: ATP Synthesis Pathway; Address; Affect; Anodes; Bacteria; Biological Assay; Biology; Bioreactors; Bioremediations; Catabolism; Cathodes; Cell membrane; Cell physiology; Cells; Chemicals; Chimeric Proteins; Communication; Couples; Cytochromes; Cytoplasm; Data; Electron Transport; Electrons; Energy-Generating Resources; Engineering; Environment; Equilibrium; Escherichia coli; Fermentation; Future; Gene Expression; Genes; Genetic; Geobacter; Growth; Heating; Investigation; Life; Light; Measures; Membrane; Membrane Proteins; Metabolism; Metals; Microbe; Microbial Biofilms; Microscopy; Modeling; Monitor; NADH; Nature; Organism; Oxidants; Oxidation-Reduction; Oxygen; Pathway interactions; Pharmacologic Substance; Photosynthesis; Potential Energy; Process; Production; Proteins; Reaction; Relative (related person); Research; Respiration; Series; Shewanella; Solid; Stimulus; System; Testing; Transcript; base; biological systems; carbon fixation; cellular engineering; clinically relevant; driving force; electrical potential; extracellular; improved; member; metabolic engineering; microbial; microorganism; mutant; pathogen; periplasm; positional cloning; public health relevance; reconstitution; respiratory; sensor; transcriptome sequencing; uptake

DESCRIPTION (provided by applicant): Living cells gain energy for growth and survival by transferring electrons between the products of catabolism and a suitable electron acceptor. The most favorable terminal electron acceptor is oxygen, but in oxygen-poor environments most cells use other soluble factors to accept the electrons generated by metabolism. Bacteria in the genus Shewanella can actually use solid extracellular metals as substrates for respiration, and they have evolved a series of periplasmic and outer-membrane proteins to serve as a direct electrical junction with these metals. Several members of the shewanellae have clinical relevance as opportunistic pathogens, and the basis of their unique respiratory abilities thus warrants further investigation. Furthermore, the existence of a genetically-encoded pathway between manufactured circuits and the central metabolism of living cells opens new vistas in studying and engineering microorganisms. Recent results suggest that these conduits can also pass current in reverse, allowing extracellular electrons to enter the cell and drive synthetic processes. If combined with carbon fixation, applied current could make a versatile input for the production of high-value chemicals, including pharmaceuticals. This proposal outlines research to characterize and improve the capacity of Shewanella cells to accept electrical current, using electrically active bioreactors in combination with microscopy of cells expressing fluorescent protein sensors, transcriptome sequencing, and reverse genetics assays. Finally, a strain of E. coli expressing genes from Shewanella will be constructed in order to define an efficient genetic circuit conferring electron uptake.

描述（由申请人提供）：活细胞通过在分解代谢产物和合适的电子受体之间转移电子来获得生长和存活能量。最有利的末端电子受体是氧气，但是在耗氧环境中，大多数细胞使用其他可溶性因子接受代谢产生的电子。 Shewanella属中的细菌实际上可以使用固体细胞外金属作为呼吸的底物，并且它们已经进化出一系列的前质和外膜蛋白，以作为与这些金属的直接电交界。 Shewanellae的几名成员作为机会病原体具有临床意义，其独特的呼吸能力的基础值得进一步研究。此外，在制造电路和活细胞的中央代谢之间存在遗传编码的途径，在研究和工程微生物中打开了新的远景。最近的结果表明，这些导管也可以反向传递电流，从而使细胞外电子进入细胞并驱动合成过程。如果与碳固定结合使用，则应用电流可能会成为生产高价值化学物质（包括药物）的多功能输入。该建议概述了研究和提高Shewanella细胞接受电流能力的研究，使用电动生物反应器与表达荧光蛋白传感器的细胞的显微镜结合使用，转录组测序和反向遗传学分析。最后，将构建来自Shewanella的大肠杆菌表达基因的菌株，以定义有效的遗传回路赋予电子摄取。

项目成果

Streptomyces thermoautotrophicus does not fix nitrogen.

• DOI: 10.1038/srep20086
• 发表时间: 2016-02-01
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: MacKellar D;Lieber L;Norman JS;Bolger A;Tobin C;Murray JW;Oksaksin M;Chang RL;Ford TJ;Nguyen PQ;Woodward J;Permingeat HR;Joshi NS;Silver PA;Usadel B;Rutherford AW;Friesen ML;Prell J
• 通讯作者: Prell J