W-Health: Tungsten is an Essential Metal for a Healthy Gut Microbiome

项目总结/摘要 NIH人类微生物组计划（HMP）彻底改变了我们对人类-微生物相互作用的看法 并为研究提供了巨大的推动力，以便更深入地了解 微生物影响人类健康。HMP参考基因组和人类的肠道微生物 胃肠道细菌培养物保藏中心包含142属961种。然而，相对而言， 对这些微生物知之甚少。在这里，我们将测试的假设，钨（W），金属 在生物系统中几乎从未被考虑过，对人类肠道微生物组的健康至关重要。我们 生物信息学分析表明，大量的这些肠道微生物含有编码不同的基因， 含钨氧化还原酶（WOR）家族的成员。只有极少数的WOR酶 已经被描述为主要来自外来的嗜热微生物。的总体目标 拟议的研究是要表明WOR家族的其他成员在肠道中具有重要功能 微生物组在初步研究中，我们已经表明，一些肠道微生物会吸收微量的W， 他们的含W的WO Rs去除肠道中发现的活性和潜在毒性的醛， 由熟食和微生物代谢产生。建议其他含W的WO R 催化其他未知的反应。在拟议的研究中，我们将纯化10个新的生物遗传学 通过W-监测（使用ICP-MS）厌氧色谱确定不同的WO。它们的催化活性和 将通过酶诱导的代谢组学方法（使用LC-MS）确定生理底物。 此外，我们提出，这些WO R中的一些是电子分支酶， 耦合放能和吸能反应，最近发现的能量守恒机制， 生物系统。该子集的动力学、光谱（使用EPR）和结构（使用cryoEM）分析 的W-酶将被用来调查的分歧反应的性质。使用基于基因组的 通过代谢重建，将确定各种WO R的生理功能，并且我们将 确定W对肠道微生物代谢的影响，包括对肠道微生物的抵抗力， 与烹饪有关的醛。 相关性：现在很清楚，除了促进消化外，肠道微生物组还在消化系统中发挥作用。 令人惊讶的广泛的人类疾病，包括帕金森氏症，精神分裂症，骨关节炎和 心血管疾病和免疫缺陷疾病。这项研究的结果将提供一个 对我们胃肠道中主要微生物的主要代谢的全新视角， 钨被认为是一种在生物系统中很少使用的金属。

Project Summary/Abstract The NIH Human Microbiome Project (HMP) revolutionized our perspective on human-microbe interactions and provided a tremendous impetus for research in order to obtain a much deeper understanding of how microbes impact human health. The gut microorganisms of the HMP Reference Genomes and the Human Gastrointestinal Bacteria Culture Collection contain 961 species representing 142 genera. Yet, relatively little is known about these microorganisms. Herein we will test the hypothesis that tungsten (W), a metal almost never considered in biological systems, is essential for the health of the human gut microbiome. Our bioinformatics analyses reveal that a large number of these gut microbes contain genes encoding diverse members of the W-containing oxidoreductase (WOR) family of enzymes. Only a very few WOR enzymes have been previously characterized, mainly from exotic thermophilic microbes. The overall goal of the proposed research is to show that other members of the WOR family have essential functions in the gut microbiome. In preliminary studies, we have shown that some gut microbes take up trace amounts of W and their W-containing WORs remove reactive and potentially toxic aldehydes found in the gut, which are generated from cooked foods and microbiome metabolism. Other W-containing WORs are proposed to catalyze other as yet unknown reactions. In the proposed research we will purify ten novel phylogenetically distinct WORs by W-monitored (using ICP-MS) anaerobic chromatography. Their catalytic activities and physiological substrates will be determined by an enzyme-induced metabolomics approach (using LC-MS). In addition, we propose that some of these WORs are electron bifurcating enzymes that simultaneously couple exergonic and endergonic reactions, a recently discovered mechanism of energy conservation in biological systems. Kinetic, spectroscopic (using EPR) and structural (using cryoEM) analyses of this subset of W-enzymes will be used to investigate the nature of the bifurcation reactions. Using genome-based metabolic reconstructions, the physiological functions of the various WORs will be ascertained and we will determine the effects of W on the metabolism of the gut microbes, including on their resistance to gut- and cooking-related aldehydes. Relevance: It is now clear that, in addition to facilitating digestion, the gut microbiome plays roles in a surprisingly extensive range of human conditions, including in Parkinson’s, schizophrenia, osteoarthritis and in cardiovascular and immune-deficiency diseases. The results of the proposed research will provide a completely new perspective on the primary metabolisms of the key microbes in our gastrointestinal tract and the proposed essential role of tungsten, a metal that was thought to be seldom used in biological systems.