权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Directed Evolution of Metal-Substituted Carbonic Anhydrases: Applications as Cat

金属取代碳酸酐酶的定向进化：作为 Cat 的应用

基本信息

批准号：
8028609
负责人：
Levi Michael Stanley
金额：
$ 8.22万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-01 至 2011-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8028609
关键词：
Active Sites Ala-Trp-Arg-His-Pro-Gln-Phe-Gly-Gly Amino Acids Area Award Azides Catalysis Chemistry Collaborations Data Development Development Plans Dialysis procedure Doctor of Philosophy Engineering Environment Enzymes Evolution Excision Exhibits Felis catus Foundations Generations Genes Goals Health Histidine Human Hydrogen Bonding Illinois Ions Isoenzymes Knowledge Laboratories Lead Libraries Mediating Medical Mentors Metals Methods Mutagenesis Mutation National Institute of General Medical Sciences National Research Service Awards Organism Pathway interactions Phase Positioning Attribute Postdoctoral Fellow Procedures Protein Engineering Protocols documentation Reaction Reporting Research Roentgen Rays Science Scientist Screening procedure Site Sulfonamides Synthesis Chemistry System Training Transition Elements Universities Zinc amidation analog base carbene carbonate dehydratase career career development catalyst cycloaddition design diazo compound directed evolution experience high throughput screening human disease improved metal complex metalloenzyme mutant nitrene novel professor programs research and development scaffold skills

项目摘要

DESCRIPTION (provided by applicant): The objectives of the research described in this Pathway to Independence Award application are to provide the candidate with training and experience in protein engineering to develop artificial metalloenzymes that catalyze new or improved synthetic reactions to prepare biologically active compounds. The candidate, Levi Stanley, Ph. D., is currently an NRSA/NIGMS postdoctoral fellow with the immediate goal of completing an additional one to two years of mentored research to establish research skills in the field of protein engineering and to apply these new skills to the development of artificial metalloenzymes that combine the broad reactivity of transition metal complexes with the ability of enzymes to exquisitely control the selectivity of synthetic transformations. Dr. Stanley's long-term career goals include establishing himself as an independent scientist in a tenure-track academic position and contributing to the improvement of human health through the development of catalytic reactions that generate bioactive compounds and building blocks to such compounds. Dr. Stanley has established expertise in transition metal catalysis through his current research in Professor John Hartwig's laboratory at the University of Illinois. The career development plan and research strategy will allow the candidate to combine his knowledge of transition metal catalysis with current methods for protein engineering. The mentored phase of the program will be carried out in collaboration with Professor John Hartwig, a leader in the field of transition metal catalysis, and Professor Huimin Zhao, an expert in the field of protein engineering. The established laboratories of the candidate's co-mentors at the University of Illinois provide an ideal environment to conduct research at the interface of their respective areas of expertise. The proposed research in the mentored phase will allow the candidate to create a novel platform for the discovery and development of new artificial metalloenzymes that catalyze reactions that are useful to synthetic chemists. The independent phase of the proposed research will focus on the application of these new artificial metalloenzymes as catalysts in regioselective, chemoselective, and stereoselective C-H functionalization, cyclopropanation, and aziridination reactions. The proposed research program is outlined in four specific aims. The first aim is focused on the identification of metal-substituted carbonic anhydrases that exhibit activity as catalysts for C-H functionalization reactions. The second aim is directed toward developing methods for the generation, expression, purification, and substitution of the metal in mutant carbonic anhydrase libraries. The third aim is designed to improve by directed evolution the activity of metal-substituted carbonic anhydrases that are active for C-H functionalization reactions. The first two aims and the beginning of the third aim will be the primary focus of the mentored (K99) phase and will provide the background necessary for the candidate to conduct protein engineering and directed evolution during the independent phase of the program. The fourth aim is directed toward the divergent evolution of metal-substituted carbonic anhydrases to identify mutants that lead to different regioselectivities and chemoselectivities from a common starting point. Research toward the completion of aims three and four will be conducted during the independent (R00) phase of this program. The results of these studies will establish a new platform to generate and improve artificial metalloenzymes and will lay the groundwork to expand the breadth of transformations catalyzed by metalloenzymes that are useful to synthetic chemists. PUBLIC HEALTH RELEVANCE: Synthetic chemistry provides a foundation for modern medical science, and the development of new catalysts and synthetic reactions lies at the forefront of chemistry targeted at generating biologically active materials. The proposed research program is focused on the study and development of new artificial metalloenzymes that combine the broad reactivity of transition metal complexes with the exquisite control of enzymes. These new metalloenzymes will be engineered to promote synthetic reactions that will ultimately facilitate the discovery of medicinal agents for treating human disease.

描述（由申请人提供）：独立之路奖申请中描述的研究目标是为候选人提供蛋白质工程方面的培训和经验，以开发人工金属酶，催化新的或改进的合成反应，以制备生物活性化合物。候选人Levi Stanley博士目前是NRSA/NIGMS博士后，其近期目标是完成额外的一到两年的指导研究，以建立蛋白质工程领域的研究技能，并将这些新技能应用于人工金属酶的开发，将过渡金属配合物的广泛反应性与酶的能力相结合，以精细地控制合成转化的选择性。他的长期职业目标包括将自己定位为终身学术职位的独立科学家，并通过开发产生生物活性化合物的催化反应和此类化合物的构建块，为改善人类健康做出贡献。他目前在伊利诺伊大学John Hartwig教授的实验室进行研究，在过渡金属催化方面建立了专业知识。职业发展规划和研究策略将允许候选人将其过渡金属催化的知识与当前蛋白质工程的方法相结合。该项目的导师阶段将与过渡金属催化领域的领导者John Hartwig教授和蛋白质工程领域的专家赵慧敏教授合作进行。候选人的共同导师在伊利诺伊大学建立的实验室为他们各自专业领域的界面进行研究提供了理想的环境。在指导阶段提出的研究将允许候选人创建一个新的平台，用于发现和开发新的人工金属酶，催化对合成化学家有用的反应。该研究的独立阶段将重点研究这些新的人工金属酶作为催化剂在区域选择性、化学选择性和立体选择性C-H功能化、环丙化和叠氮化反应中的应用。拟议的研究计划概述为四个具体目标。第一个目标是集中在鉴定金属取代的碳酸酐酶，表现出活性作为C-H功能化反应的催化剂。第二个目标是开发突变型碳酸酐酶文库中金属的生成、表达、纯化和替代方法。第三个目标是通过定向进化来提高对碳氢官能化反应有活性的金属取代碳酸酐酶的活性。前两个目标和第三个目标的开始将是指导（K99）阶段的主要焦点，并将为候选人在项目的独立阶段进行蛋白质工程和定向进化提供必要的背景。第四个目标是针对金属取代碳酸酐酶的不同进化，从一个共同的起点识别导致不同区域选择性和化学选择性的突变体。完成目标3和目标4的研究将在该计划的独立（R00）阶段进行。这些研究结果将为合成和改进人工金属酶提供一个新的平台，并为扩大金属酶催化转化的广度奠定基础。