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的看法。 微生物相互作用，并为研究提供了巨大的动力，以获得更多的 深入了解微生物如何影响人类健康。HMP的肠道微生物 参考基因组和人类胃肠道细菌培养物保藏中心包含961个 142属142种。然而，对这些微生物的了解相对较少。 在这里，我们将测试的假设，钨（W），金属几乎从来没有考虑在生物 系统，对人类肠道微生物组的健康至关重要。我们的生物信息学分析 揭示了大量的这些肠道微生物含有编码不同成员的基因， 含钨氧化还原酶（WOR）家族的酶。只有极少数WOR酶具有 以前的特点，主要是外来的嗜热微生物。的总体目标 拟议的研究是为了表明WOR家族的其他成员在以下方面具有重要功能： 肠道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 一定量的W和它们的含W的WO R除去反应性和潜在毒性的醛 在肠道中发现，这是从熟食和微生物代谢产生的。其他W- 含有WO R的催化剂被提议用于催化其它未知的反应。拟议 我们将通过W-监测（使用ICP-MS）纯化十种新的遗传学上不同的WO R。 厌氧层析它们的催化活性和生理底物将是 通过酶诱导代谢组学方法（使用LC-MS）测定。另外我们 我认为这些WO R中的一些是电子分叉酶， 放能和吸能反应，最近发现的能量守恒机制， 生物系统。的动力学、光谱（使用EPR）和结构（使用cryoEM）分析。 W-酶的这一亚类将用于研究分叉反应的性质。使用 基于基因组的代谢重建，各种WO R的生理功能将 我们将确定W对肠道微生物代谢的影响， 包括它们对肠道和烹饪相关醛的抗性。

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.