Collaborative Research: Ethylene-Forming Enzyme

合作研究：乙烯形成酶

• 批准号: 1904215
• 负责人: Christo Christov
• 金额: $ 17.9万
• 依托单位: Michigan Technological University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1904215&HistoricalAwards=false
• 关键词: Collaborative; Research; Ethylene; Forming; Enzyme

Guanidine is a potential nitrogen fertilizer and ethylene has been proposed as a potential alternative fuel source; thus, the biosynthetic reactions that form these compounds are commercially important as they have potential benefits for agriculture and the biofuels industries. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Drs. Robert Hausinger and Jian Hu at Michigan State University, and Dr. Christo Christov at Michigan Technological Institute to investigate how ethylene-forming enzyme (EFE) from bacteria and fungi catalyzes dual reactions to form guanidine or ethylene. In the typical reaction of EFE, a carbon dioxide is removed from other molecules (2-oxoglutarate, or 2OG), while being added to the amino acid, L-arginine, to transform it into guanidine. Alternatively, EFE catalyzes the transformation of 2OG into ethylene, with the release of carbon dioxide / bicarbonate. Research into the mechanisms of EFE catalysis provides specialized biochemical, structural, and computational training to postdoctoral fellows, and graduate and undergraduate students, including women and students from underrepresented minority groups. Incorporation of these studies into a university science festival engages families and lifelong learners through outreach programs about the potential use of ethylene as a biofuel.The overarching goals of this research project are to elucidate the molecular determinants that distinguish the two distinct reactions catalyzed by EFE and to establish the catalytic mechanisms of the enzyme by a combination of experimental and computational studies. This work also investigates why other members of the iron(II)- and 2OG-dependent oxygenases fail to produce ethylene. To understand the basis of the two EFE-catalyzed reactions, the researchers are characterizing naturally occurring analogs, a reconstructed ancestor, and site-directed variants of the protein by using kinetic, structural, and computational methods. Using this information along with structure-guided protein engineering, the researchers are creating EFE variants optimized for the production of either the biofuel ethylene or the plant fertilizer guanidine while eliminating wasteful L-Arginine hydroxylation or 2OG-degrading reactions. In addition, the team is using biochemical, kinetic, spectroscopic, and computational methods to elucidate the chemical reaction mechanism of EFE. This work extends the structural/mechanistic information available on other important iron(II)- and 2OG-dependent oxygenases that are unable to catalyze these reactions.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

胍是一种潜在的氮肥，乙烯已被提议作为潜在的替代燃料来源;因此，形成这些化合物的生物合成反应具有商业重要性，因为它们对农业和生物燃料行业具有潜在的好处。有了这个奖项，化学部的生命过程化学计划正在资助密歇根州立大学的Robert Hausinger和Jian Hu博士以及密歇根技术研究所的Christo Christov博士研究来自细菌和真菌的乙烯形成酶（EFE）如何催化双重反应形成胍或乙烯。在EFE的典型反应中，二氧化碳从其他分子（2-酮戊二酸或2 OG）中除去，同时加入氨基酸L-精氨酸，将其转化为胍。 或者，EFE催化2 OG转化为乙烯，释放二氧化碳/碳酸氢盐。EFE催化机制的研究为博士后研究员，研究生和本科生，包括妇女和代表性不足的少数群体的学生提供专业的生物化学，结构和计算培训。将这些研究纳入大学科学节，通过关于乙烯作为生物燃料的潜在用途的推广计划吸引家庭和终身学习者。该研究项目的总体目标是阐明区分EFE催化的两种不同反应的分子决定因素，并通过实验和计算研究的结合建立酶的催化机制。这项工作还调查了为什么其他成员的铁（II）和2 OG依赖性加氧酶不能产生乙烯。为了了解这两个EFE催化反应的基础，研究人员正在通过使用动力学，结构和计算方法来表征天然存在的类似物，重建的祖先和蛋白质的定点变体。利用这些信息沿着结构导向的蛋白质工程，研究人员正在创造EFE变体，这些变体被优化用于生产生物燃料乙烯或植物肥料胍，同时消除浪费的L-精氨酸羟基化或2 OG降解反应。此外，该团队正在使用生物化学，动力学，光谱和计算方法来阐明EFE的化学反应机制。这项工作扩展了其他重要的铁（II）和2 OG依赖的加氧酶，不能催化这些reactions.This奖项反映了NSF的法定使命的结构/机制信息，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

项目成果

Dioxygen Binding Is Controlled by the Protein Environment in Non‐heme Fe II and 2‐Oxoglutarate Oxygenases: A Study on Histone Demethylase PHF8 and an Ethylene‐Forming Enzyme

非血红素 Fe II 和 2-氧化戊二酸加氧酶中的双氧结合受蛋白质环境控制：组蛋白脱甲基酶 PHF8 和乙烯形成酶的研究

• DOI: 10.1002/chem.202300138
• 发表时间: 2023
• 期刊: Chemistry – A European Journal
• 作者: Chaturvedi, Shobhit S.;Thomas, Midhun George;Rifayee, Simahudeen Bathir Jaber Sathik;White, Walter;Wildey, Jon;Warner, Cait;Schofield, Christopher J.;Hu, Jian;Hausinger, Robert P.;Karabencheva‐Christova, Tatayana G.
• 通讯作者: Karabencheva‐Christova, Tatayana G.

Catalytic Mechanism of Human Ten-Eleven Translocation-2 (TET2) Enzyme: Effects of Conformational Changes, Electric Field, and Mutations

• DOI: 10.1021/acscatal.0c05034
• 发表时间: 2021-03-14
• 期刊: ACS CATALYSIS
• 影响因子: 12.9
• 作者: Waheed, Sodiq O.;Chaturvedi, Shobhit S.;Christov, Christo Z.
• 通讯作者: Christov, Christo Z.

Role of Structural Dynamics in Selectivity and Mechanism of Non-heme Fe(II) and 2-Oxoglutarate-Dependent Oxygenases Involved in DNA Repair

• DOI: 10.1021/acscentsci.0c00312
• 发表时间: 2020-05-27
• 期刊: ACS CENTRAL SCIENCE
• 影响因子: 18.2
• 作者: Waheed, Sodiq O.;Ramanan, Rajeev;Karabencheva-Christova, Tatyana G.
• 通讯作者: Karabencheva-Christova, Tatyana G.

Catalysis by the Non-Heme Iron(II) Histone Demethylase PHF8 Involves Iron Center Rearrangement and Conformational Modulation of Substrate Orientation

• DOI: 10.1021/acscatal.9b04907
• 发表时间: 2020-01-17
• 期刊: ACS CATALYSIS
• 影响因子: 12.9
• 作者: Chaturvedi, Shobhit S.;Ramanan, Rajeev;Christov, Christo Z.
• 通讯作者: Christov, Christo Z.