Biosynthesis of the Iron-Molybdenum Cofactor

铁钼辅因子的生物合成

• 批准号: 0920380
• 负责人: Robert Igarashi
• 金额: $ 52.05万
• 依托单位: The University of Central Florida Board of Trustees
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0920380&HistoricalAwards=false
• 关键词: Biosynthesis; Iron; Molybdenum; Cofactor

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).Intellectual Merit The iron-molybdenum cofactor (FeMo-co, 7Fe-9S-Mo-X-homocitrate), found within the active site of the enzyme nitrogenase, is among nature's most complex hetero-metallic cofactors and is required to catalyze the chemically difficult reduction of dinitrogen (N2) to ammonia (NH3). FeMo-co is synthesized by a multi-step pathway with the involvement of multiple different proteins that couple Fe, S and Mo. Currently, the exact structures of the pathway intermediate Fe-S clusters are unknown and even less is known regarding the molecular mechanisms of the enzymes such as NifB and NifEN that catalyze the bio-metallic reactions. By combined genetic, biochemical and spectroscopic techniques, this research aims to determine the exact structures of the pathway intermediates and uncover the enzymatic mechanisms that perform the multi-step bio-metallic reactions that are the foundation of biological nitrogen fixation. The major focus will be placed on NifB and its product, NifB-co, that form the early part of the FeMo-co biosynthetic pathway. The NifB enzyme, NifB-co, as well as isotopic analogs, will be examined by EPR, Mössbauer, EXAFS and NRVS spectroscopic techniques. These spectroscopic inspections will be complemented by biochemical analysis to determine the exact step(s) and mode of incorporation of each Fe, S and the interstitial atom X. Broader ImpactAll life depends on the input of the element nitrogen into the biosphere by biological nitrogen fixation catalyzed by FeMo-co. Understanding how the enzymes of the FeMo-co biosynthesis pathway perform the bio-metallic chemistry may lead to design and synthesis of novel catalysts to improve Haber-Bosch processes for fertilizer production resulting in a decrease in energy costs for agriculture. Both graduate and undergraduate students will have a major role in this research and will be trained in a wide range of cross-disciplinary techniques. As a part of this project, an outreach activity that targets junior and senior High School students will be performed. The designed activity will bring biochemistry into the secondary education classroom.

该奖项由2009年美国复苏和再投资法案（Public Law 111-5）资助。知识产权在固氮酶的活性部位发现的铁钼辅因子（FeMo-co，7 Fe-9 S-Mo-X-homocitrate）是自然界最复杂的异金属辅因子之一，需要催化化学上困难的二氮（N2）还原为氨（NH3）。 FeMo-co是通过多步途径合成的，其中涉及多种不同的蛋白质，这些蛋白质将Fe，S和Mo偶联。 目前，途径中间体Fe-S簇合物的确切结构是未知的，关于催化生物金属反应的酶如NifB和NifEN的分子机制更是知之甚少。 本研究旨在通过结合遗传学、生物化学和光谱技术，确定途径中间体的确切结构，并揭示进行多步生物金属反应的酶机制，这些反应是生物固氮的基础。 主要重点将放在NifB及其产物NifB-co上，它们构成FeMo-co生物合成途径的早期部分。 NifB酶，NifB-co，以及同位素类似物，将通过EPR，穆斯堡尔，EXAFS和NRVS光谱技术进行检查。 这些光谱检查将通过生化分析进行补充，以确定每个Fe、S和间隙原子X的确切步骤和结合模式。 更广泛的影响所有的生命都依赖于通过FeMo-co催化的生物固氮将元素氮输入生物圈。了解FeMo-co生物合成途径的酶如何执行生物金属化学可能会导致设计和合成新型催化剂，以改善肥料生产的Haber-Bosch过程，从而降低农业的能源成本。 研究生和本科生都将在这项研究中发挥重要作用，并将接受广泛的跨学科技术培训。 作为该项目的一部分，将开展一项针对初中和高中学生的外联活动。设计的活动将把生物化学带入中学教育课堂。