Mechanistic Studies on Fe-Type Nitrile Hydration Catalysts

Fe型腈水合催化剂的机理研究

• 批准号: 1808711
• 负责人: Brian Bennett
• 金额: $ 52.5万
• 依托单位: Marquette University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1808711&HistoricalAwards=false
• 关键词: Mechanistic; Studies; Fe; Type; Nitrile

This award by the Chemistry of Life Processes in the Chemistry Division, is funding Drs. Bennett and Holz from Marquette University to investigate how the enzyme Nitrile hydratase (NHase) transforms chemical groups called nitriles into amides under ambient conditions. Interestingly, NHases can hydrate not only nitriles found in biological systems but also a wide range of synthetic nitriles. Hence NHases are extensively used in preparative organic chemistry laboratories and in the industrial production of acrylamide and nicotinamide. NHases have also proven useful in the cleanup of nitrile-based chemicals and pesticides from the environment and are thus becoming increasingly recognized as a new type of "Green" catalyst. Knowledge of the mechanisms that underscore their chemical reactivity and applications could provide an impetus for the development of new biocatalysts. Drs. Bennett and Holz's research use an interdisciplinary experimental approach to gain insight into how nitriles are hydrated by NHases. Their research and education efforts help promote teaching, training and learning for graduate and undergraduate students, particularly those from underrepresented groups. Their research also enhances the infrastructure for research and education at Marquette University. While the biological roles of NHases are not well understood, they likely include nutrient metabolism, nitrile detoxification and/or nutrient assimilation. Nitriles are most commonly found in biological systems as cyanoglycosides or cyanolipids, but many plants also produce nitriles such as ricin and beta-cyanoalanine for self-defense purposes. The major impediment to understanding their biological and bioremediation roles is the lack of understanding of their catalytic mechanism. Drs. Bennett and Holz provide molecular level insight into the hydration of nitriles by NHases using a combination of kinetics, spectroscopy, biochemistry, computational chemistry, and X-ray crystallography studies. Answers to three specific questions are sought: (1) What is the transition-state of nitrile hydration? (2) Is a transient disulfide bond between the sulfenic acid cyclic intermediate and the axial alpha-Cys ligand required for catalysis? and (3) What controls metallocenter assembly in Fe-type NHases? Successful completion of this project enhances our understanding of the catalytic mechanism of NHases and stimulate development of engineered bioremediation catalysts.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.

化学过程中化学过程的奖项是资助Drs的资金。 Marquette大学的Bennett和Holz研究氮化酶（NHASE）如何在环境条件下将称为硝酸盐的化学基团转化为酰胺。 有趣的是，NHASE不仅可以在生物系统中发现的硝酸盐，而且可以补充各种合成硝化液。 因此，NHase广泛用于制备有机化学实验室以及丙烯酰胺和烟酰胺的工业生产中。 事实证明，NHases在清理基于硝酸盐的化学物质和农药中有用，因此越来越被公认为是一种新型的“绿色”催化剂。 了解强调其化学反应性和应用的机制可以为开发新的生物催化剂提供动力。 博士。贝内特（Bennett）和霍尔兹（Holz）的研究采用跨学科的实验方法来深入了解硝酸盐如何通过NHASE水化。 他们的研究和教育工作有助于促进研究生和本科生的教学，培训和学习，尤其是来自代表性不足的人的教学和学习。 他们的研究还增强了马奎特大学的研究和教育基础设施。 虽然NHases的生物学作用尚不清楚，但它们可能包括营养代谢，硝酸盐排毒和/或营养同化。 硝酸盐最常见于生物系统中，作为氰基糖苷或氰基脂质，但许多植物也生产硝酸盐，例如ricin和beta-cyanoalanine，用于自卫目的。 了解其生物学和生物修复作用的主要障碍是对其催化机制的理解不足。 博士。 Bennett和Holz通过动力学，光谱，生物化学，计算化学和X射线晶体学研究的组合，通过NHASE提供了分子水平的洞察力。 寻求三个特定问题的答案：（1）硝酸盐水合的过渡状态是什么？ （2）在催化中所需的硫酸环状中间体和轴向α-cys配体之间的瞬时二硫键？ （3）什么控制了Fe-Type Nhass中的金属中心组件？ 该项目的成功完成增强了我们对NHASE催化机制的理解，并刺激了工程生物修复催化剂的发展。该奖项反映了NSF的法定任务，并被认为是通过基金会的智力优点和更广泛的影响来通过评估来获得支持的。

项目成果

Insights into the catalytic mechanism of a bacterial hydrolytic dehalogenase that degrades the fungicide chlorothalonil

• DOI: 10.1074/jbc.ra119.009094
• 发表时间: 2019-07
• 期刊: The Journal of Biological Chemistry
• 作者: Xinhang Yang;B. Bennett;R. Holz

Structural basis for the hydrolytic dehalogenation of the fungicide chlorothalonil

• DOI: 10.1074/jbc.ra120.013150
• 发表时间: 2020-06-26
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Catlin，Daniel S.;Yang，Xinhang;Liu，Dali
• 通讯作者: Liu，Dali

The Fe-type nitrile hydratase from Rhodococcus equi TG328-2 forms an alpha-activator protein complex

马红球菌 TG328-2 的 Fe 型腈水合酶形成 α 激活蛋白复合物

• DOI: 10.1007/s00775-020-01806-y
• 发表时间: 2020
• 期刊: JBIC Journal of Biological Inorganic Chemistry
• 作者: Lankathilaka, K. P.;Bennett, Brian;Holz, Richard C.
• 通讯作者: Holz, Richard C.

Cellular maturation of an iron-type nitrile hydratase interrogated using EPR spectroscopy

• DOI: 10.1007/s00775-019-01720-y
• 发表时间: 2019-10-01
• 期刊: JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY
• 影响因子: 3
• 作者: Lankathilaka, K. P. Wasantha;Stein, Natalia;Bennett, Brian
• 通讯作者: Bennett, Brian

Examination of the Catalytic Role of the Axial Cystine Ligand in the Co-Type Nitrile Hydratase from Pseudonocardia thermophila JCM 3095

轴向胱氨酸配体在嗜热假诺卡氏菌 JCM 3095 共型腈水合酶中的催化作用的检查

• DOI: 10.3390/catal11111381
• 发表时间: 2021
• 期刊: Catalysts
• 影响因子: 3.9
• 作者: Ogutu, Irene R.;St. Maurice, Martin;Bennett, Brian;Holz, Richard C.
• 通讯作者: Holz, Richard C.