Collaborative Research: Mechanistic Studies on Fe-type Nitrile Hydration Catalysts

合作研究：铁型腈水合催化剂的机理研究

• 批准号: 1462201
• 负责人: Brian Bennett
• 金额: $ 10.2万
• 依托单位: Marquette University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1462201&HistoricalAwards=false
• 关键词: Collaborative; Research; Mechanistic; Studies; Fe

Nitriles are used extensively in the production of specialty chemicals like the polyamide nylon. Nitrile hydratases (NHases) catalyze the hydration of nitriles to amides at ambient temperatures in neutral water. The currently employed industrial conditions for the hydration of nitriles to amides use either strong acids or bases, and are often incompatible with the sensitive structures of the compounds to be produced. NHases have therefore attracted substantial interest as biocatalysts in preparative organic chemistry. Despite the industrial importance of NHase enzymes, the details of how they work remain poorly understood. The objective of this research proposal is to fill this knowledge gap. The interdisciplinary nature of the effort will serve as a platform to increase the participation of women and classically underrepresented graduate and undergraduate students in scientific careers in biophysics, chemistry, and biochemistry. As part of the proposed project, both PI's will participate in the SMART program that mentors Milwaukee area public high school students in research projects.With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Drs. Richard C. Holz and Brian Bennett from Marquette University to elucidate the catalytic mechanism of NHase enzymes. An interdisciplinary approach will be used that will incorporate kinetics, spectroscopy, biochemistry, computational chemistry, and both X-ray and neutron crystallography. The proposed experimental approach is designed to address three key questions: i) What is the transition-state structure of nitrile hydration? ii) What are the protonation states of catalytically important species throughout the reaction? and iii) What controls metallocenter assembly in a purported eukaryotic NHase? The successful completion of this research will greatly benefit society by facilitating the intelligent design and manufacture of nitrile-based chiral pharmaceuticals and industrially important specialty chemicals, with enhanced synthetic flexibility and lower environmental impact, than at present.

腈广泛用于生产特种化学品，如聚酰胺尼龙。 腈水合酶（NHases）在环境温度下在中性水中催化腈水合为酰胺。 目前用于腈水合成酰胺的工业条件使用强酸或强碱，并且通常与待生产的化合物的敏感结构不相容。因此，作为制备有机化学中的生物催化剂，NHases引起了人们极大的兴趣。 尽管腈水合酶的工业重要性，他们如何工作的细节仍然知之甚少。 本研究的目的是填补这一知识空白。 这项工作的跨学科性质将作为一个平台，以增加妇女和传统上代表性不足的研究生和本科生在生物物理学，化学和生物化学的科学生涯的参与。 作为拟议项目的一部分，两位PI将参加SMART计划，该计划指导密尔沃基地区的公立高中学生进行研究项目。Holz和来自马奎特大学的Brian班尼特阐明了腈水合酶的催化机制。 一个跨学科的方法将被使用，将结合动力学，光谱学，生物化学，计算化学，X射线和中子晶体学。 所提出的实验方法旨在解决三个关键问题：i）腈水合的过渡态结构是什么？ ii）整个反应过程中催化重要物质的质子化状态是什么？和iii）在所谓的真核生物的NHase中，是什么控制着转录中心的组装？ 这项研究的成功完成将大大有利于社会，促进智能设计和制造腈基手性药物和工业上重要的特种化学品，提高合成灵活性和降低环境影响，比现在。

项目成果

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