De novo designed metalloproteins as a new generation of artificial hydrogenases

从头设计金属蛋白作为新一代人工氢化酶

基本信息

批准号：
10360279
负责人：
Saumen Chakraborty
金额：
$ 41.95万
依托单位：
UNIVERSITY OF MISSISSIPPI
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-20 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10360279
关键词：
Active Sites Affect Bioinorganic Chemistry Biotechnology Carbon Catalysis Chemistry Complex Dependence Development Electrochemistry Electron Transport Environment Enzymes Film Fluorescence Future Gases Generations Goals Hydrogen Bonding Hydrogenase Kinetics Location Mentors Metal Binding Site Metalloproteins Metals Methods Mississippi Modeling Mononuclear Multienzyme Complexes Natural Resources Organism Outcome Outcome Study Oxidation-Reduction Pathway interactions Peptides Phase Photosensitizing Agents Production Property Protein Engineering Proteins Reaction Research Research Activity Role Scaffolding Protein Science Security Site Solid Source Spectroscopy, Fourier Transform Infrared Spectrum Analysis Structure Sulfhydryl Compounds Sunlight Surface System Testing Titrations Training Universities Water Work absorption analog base biophysical techniques carbon emissions career catalyst cofactor design dimer finesse graduate student improved functioning insight metallicity metalloenzyme nickel-iron hydrogenase novel programs scaffold small molecule synergism undergraduate student

项目摘要

PROJECT SUMMARY/ABSTRACT Hydrogenases are complex, metal-containing enzymes that generate energy for certain organisms by catalyzing the reversible interconversion between H+ and H2 gas. Unraveling the intricate details about the function of these enzymes will significantly advance the H2-based, carbon-neutral alternative energy production. However, the complexity of these enzymes due to the presence of multiple metallic cofactors, low production yield, and deactivation, makes studying these enzymes challenging. Our long term goals are to design artificial biomolecular hydrogenases (ArHs) as simpler functional analogs of these metalloenzymes. De novo metalloprotein design is an appealing and well-established approach to model complex metalloproteins within minimal protein scaffolds. Although the designed systems are less complex, they serve as water-soluble functional analogs of the native metalloenzymes and provide a functional view of the chemistry. Employing this approach, we propose to pursue three Specific Aims describing the overall design principles and functional/mechanistic attributes of the ArHs inspired by the [NiFe] hydrogenases. The overall objectives of this proposal are: i) to design mononuclear (Ni), binuclear (Ni-Fe), and multinuclear (Ni3) active sites within suitable de novo scaffolds; ii) characterize the physical and catalytic properties of the ArHs; iii) determine the timescales of electron transfer; iv) outline the H+ transfer pathways; v) characterize the reaction intermediates; and vi) elucidate how metals and protein scaffold work in synergy to influence the properties/reactivity, such that a holistic mechanistic view of H-H bond formation can be attained. Our strong preliminary results presented here attest that our objectives are achievable. Collectively, the results from this proposed work will impact the fields of metalloprotein design, bioinorganic chemistry, and alternative energy research. A novel class of ArHs will emerge, which will provide functional vignettes into the working principles of H+ reduction pertaining to the native enzymes. The modular design parameters and outcomes from this study will enable us to prepare biosynthetic catalysts with novel properties and functions in the future.

项目摘要/摘要氢化酶是复杂的，具有金属的酶，可通过催化H+和H2气体之间的可逆互连。揭开有关的复杂细节这些酶的功能将显着提高基于H2的碳中性替代能生产。但是，由于存在多个金属辅因子，这些酶的复杂性，低生产产量和失活使研究这些酶具有挑战性。我们的长期目标是将人工生物分子氢化酶（ARHS）设计为这些金属酶的更简单的功能类似物。从头开始金属蛋白设计是一种吸引人且良好的模型复杂方法最小蛋白支架内的金属蛋白。尽管设计的系统不那么复杂，但它们服务作为天然金属酶的水溶性功能类似物，并提供了功能视图化学。采用这种方法，我们建议追求描述整体设计的三个特定目标受[Nife]氢化酶启发的ARH的原理和功能/机械属性。总体该建议的目标是：i）设计单核（NI），双核（NI-FE）和多核（NI3）Active 合适的从头脚手架中的地点； ii）表征ARH的物理和催化特性； iii）确定电子传输的时间尺度； iv）概述H+转移途径； v）表征反应中间人； vi）阐明金属和蛋白质支架如何在协同作用中起作用以影响属性/反应性，使得可以实现H-H键形成的整体机械视图。我们的坚强这里提出的初步结果证明我们的目标是可以实现的。总的来说，这项拟议工作的结果将影响金属蛋白设计，生物无机的领域化学和替代能源研究。一类新颖的ARHS将出现，该类别将提供功能插入与天然酶有关的H+还原的工作原理。模块化设计这项研究的参数和结果将使我们能够制备具有新型特性的生物合成催化剂和将来的功能。