Discovery and application of new halogenases

新型卤化酶的发现及应用

• 批准号: 10392879
• 负责人: MICHELLE C CHANG
• 金额: $ 31.46万
• 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 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392879
• 关键词: Active Sites; Alkynes; Amino Acids; Anabolism; Anions; Binding Proteins; Biochemical; Bioinformatics; Biological; Biology; Carbon; Catalysis; Cells; Chemicals; Chemistry; Complex; Development; Electrons; Engineering; Enzymes; Evolution; Face; Family; Family member; Foundations; Generations; Geometry; Goals; Halogens; Health; Human; Hydrogen Bonding; Hydroxylation; In Vitro; Indole Alkaloids; Libraries; Link; Metabolic Biotransformation; Methodology; Modification; Molecular; Natural Products; Nature; Outcome; Pathway interactions; Peptides; Positioning Attribute; Production; Protein Engineering; Proteins; Reaction; Ribosomes; Route; Source; Structure; Translations; base; catalyst; chemical function; design; functional group; halogenation; improved; in vivo; innovation; insight; interest; novel; public health relevance; restraint; scaffold; screening; small molecule; tool

Project Summary/Abstract 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, which remains challenging to achieve. 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, and (iii) developing applications for halogenases to produce new amino-acid based products.

项目摘要/摘要 分子多样性的快速和模块化产生是寻找新化学化学物质的关键 功能。一个特别有用的功能组是卤素（x = cl，br，i），它可以实现 许多选择性有效的下游策略来创造结构复杂性。在这个 考虑到卤代酶为 用于区域和立体选择性引入卤素取代基的合成催化剂 复杂的脚手架，这仍然具有挑战性。而许多卤素酶家族 存在，自由基卤素酶为反应多样性提供了最大的潜力，因为它们是 有能力用卤素代替未激活的C-H键 亲电或亲核机制。但是，这些酶的底物范围具有 迄今为止限制到蛋白质结合的底物或大型晚期天然产品 中间人。我们的小组发现了一个新的自由基卤素酶，能够反应 与小分子底物。我们现在寻求利用这一发现来开发新的 体外和体内合成的工具。该提案的具体目的包括：（i）阐明 这些新的自由基卤素酶的结构和机制，这将提供重要的见解 进入他们的工程，（ii）调查和工程选择性的卤素化合物，以及（iii） 开发用于产生新的基于氨基酸产品的卤代酶的应用。