Collaborative Research: The Membrane Associated Methane Monooxygenase from Methylococcus capsulatus Bath: Metal Centers and Mechanism of Substrate Hydroxylation

NSF化学部生命过程化学(CLP)项目的这一奖项支持爱荷华州立大学的Alan A.DiSpirito教授和卡内基梅隆大学的Eckard Munck教授的一个合作项目，该项目旨在开展基础研究，旨在阐明活性中心的结构和所谓的颗粒形式甲烷单加氧酶(PMMO)的反应性。这是一种普遍存在的细菌酶，介导甲烷转化为甲醇，因此在全球碳循环中发挥关键作用。在温和的反应条件下，甲烷选择氧化制甲醇是金属有机化学家追求的最重要的工业过程之一，可以直接应用于pMMO活性中心的结构解析。该项目包括广泛使用穆斯堡尔谱来表征pMMO中所谓的双铁中心，使用电子顺磁共振(EPR)光谱识别催化和电子转移中心，以及甲烷氧化的机理研究。这是一个涉及生物化学、微生物生理学和生物物理化学的卓越的跨学科项目，将使本科生和研究生接触到从蛋白质纯化到EPR光谱分析的各种技术。该提案的更广泛影响集中在学生的培训上，包括来自代表性不足群体的成员和威斯康星大学Eau Claire大学的本科生，该大学是一个以本科生为主的机构。

This award in the Chemistry of Life Processes (CLP) program in the Division of Chemistry at NSF supports a collaborative project between Professor Alan A. DiSpirito at Iowa State University and Professor Eckard Munck at Carnegie Mellon University to carry out fundamental studies aimed at elucidating the structure of the active site and the reactivity of the so-called particulate form of methane monooxygenase (pMMO). This is a ubiquitous bacterial enzyme that mediates the conversion of methane to methanol and thus plays a key role in the global carbon cycle. The selective oxidation of methane to methanol using mild reaction conditions is one of the most important industrial processes pursued by organometallic chemists and could be a direct application of the structural elucidation of pMMO's active site. The project includes the extensive use of Mössbauer spectroscopy to characterize the purported diiron center in pMMO, the identification of catalytic and electron transfer centers using Electron Paramagnetic Resonance (EPR) spectroscopy, and a mechanistic investigation of methane oxidation. This is an eminently interdisciplinary project at the interface of biochemistry, microbial physiology, and biophysical chemistry and will expose undergraduate and graduate students to a variety of techniques, ranging from protein purification to EPR spectroscopy.The broader impacts of the proposal are focused on the training of students, including members from underrepresented groups and undergraduate students from the University of Wisconsin-Eau Claire, a Predominantly Undergraduate Institution.