Discovery and Application of New Halogenases

新型卤化酶的发现及应用

• 批准号: 10798668
• 负责人: MICHELLE C CHANG
• 金额: $ 6.46万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10798668
• 关键词: Amino Acids; Analytical Chemistry; Binding Proteins; Calibration; Carbon; Charge; Chemistry; Chromatography; Complex; Continuity of Patient Care; Cyclization; Data; Dedications; Distant; Engineering; Enzymes; Equipment; Family; Funding; Generations; Halogens; High Pressure Liquid Chromatography; Hour; Hydrogen Bonding; In Vitro; Injections; Investments; Laboratories; Libraries; Maintenance; Modification; Molecular; Natural Products; Notification; Performance; Positioning Attribute; Process; Protein Engineering; Proteins; Pump; Reaction; Research; Running; Sampling; Schedule; Services; Structure; Students; System; Training; Variant; Water; Work; carbene; catalyst; chemical function; detector; end of life; fast protein liquid chromatography; functional group; halogenation; in vivo; insight; instrument; member; mutant; professor; scaffold; scale up; small molecule; stereochemistry; timeline; tool

Project Summary The rapid and modular generation of molecular diversity is key to the search for new chemical functions. One particularly useful functional group is the halogen (X = Cl, Br, I), which enables many selective and effective downstream strategies for creating structural complexity. In this regard, halogenase enzymes have provided an important and complementary approach to synthetic catalysts for regio- and stereoselective introduction of a halogen substituent on a complex scaffold. While many families of halogenases exist, the radical halogenases provide the greatest potential for reaction diversity, as they are competent to replace unactivated C-H bonds with a halogen unlike those that operate by electrophilic or nucleophilic mechanisms. However, the substrate scope of these enzymes has been limited to date to either protein-bound substrates or large late-stage natural product intermediates. Our group has discovered a new clade of radical halogenases capable of reacting with small molecule substrates. We now seek to take advantage of this discovery to develop new tools for in vitro and in vivo synthesis. Specific aims of this proposal include: (i) elucidating the structure and mechanism of these new radical halogenases, which will provide important insight into their engineering, (ii) investigating and engineering selectivity in halogenases.

项目摘要 分子多样性的快速和模块化生成是寻找新化学品的关键 功能协调发展的一种特别有用的官能团是卤素（X = Cl、Br、I），其使得许多 创造结构复杂性的选择性和有效的下游战略。在这方面，卤化酶 酶提供了一种重要的和补充的方法来合成催化剂， 在复杂支架上立体选择性引入卤素取代基。虽然许多家庭 如果存在卤化酶，则自由基卤化酶提供反应多样性的最大潜力，因为它们是 能够用卤素取代未活化的C-H键，不像那些通过亲电或 亲核机制然而，迄今为止，这些酶的底物范围仅限于 蛋白质结合的底物或大的后期天然产物中间体。我们小组发现了一种新的 能够与小分子底物反应的自由基卤化酶的进化枝。我们现在寻求 这一发现的优势是开发用于体外和体内合成的新工具。具体目标 建议包括：（i）阐明这些新的自由基卤代酶的结构和机制，这将 提供重要的洞察他们的工程，（二）调查和工程选择性， 卤化酶