Biosynthesis of unusual bio-orthogonal functionalities in natural products

天然产物中不寻常的生物正交功能的生物合成

• 批准号: 10600347
• 负责人: Wenjun Zhang
• 金额: $ 7.9万
• 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: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10600347
• 关键词: Alkynes; Anabolism; Biochemical; Biological; Biology; Biotechnology; C-terminal; Carboxy-Lyases; Chemicals; Enzymatic Biochemistry; Enzymes; Generations; Goals; Health; Human; Interdisciplinary Study; Iron; Logic; Membrane; Methionine-tRNA Ligase; N Domain; N-terminal; Natural Products; Oxidases; Parents; Pathway interactions; Personal Satisfaction; Public Health; Research; System; alpha ketoglutarate; bioactive natural products; chemical property; desaturase; design; enzyme pathway; flexibility; functional group; heme a; human disease; innovation; metalloenzyme; novel; pharmacophore; physical property; public health relevance; small molecule; tool

SUMMARY Natural products are important small molecules for studying, treating, and even causing human diseases, and they typically have unique functional groups that are critical for their biological activities. By exploiting the biosynthetic machinery by which these functionalities are synthesized, it is possible to enhance, vary or diminish the biological activities of parent compounds and apply the biosynthetic machinery to new systems for functional group installation. Toward this goal, the chemical logic and enzymatic machinery underlying natural product biosynthesis need to be fully characterized and understood. This project will focus on functional characterization and mechanistical interrogation of enzymes responsible for biosynthesis of unusual pharmacophores of natural products, including terminal alkyne, isonitrile and N-hydroxytriazene. Unique metalloenzymes have been identified important for their biosynthesis, including the membrane-bound di-iron dependent bifunctional desaturase/acetylenase for alkyne generation, the non-heme iron and α-ketoglutarate dependent oxidase/decarboxylase for isonitrile synthesis, and a didomain enzyme consisting of a C-terminal methionyl- tRNA synthetase-like domain and an N-terminal cupin domain for N-hydroxytriazene formation. The biochemical and mechanistic understanding of these newly discovered enzymes remain poor, warranting an in-depth study in this project. In addition to serving as the warheads of bioactive natural products, these functionalities have distinct chemical and physical properties and are utilized in bio-orthogonal chemical transformations for various chemical biology applications. We will thus further explore the application of their biosynthetic machinery to install these “clickable” functionalities on various biomolecules on demand. The research strategy is innovative in terms of the choice of novel functionalities and their unique but poorly understood biosynthetic machinery, the multidisciplinary research tools, and the goal-oriented approaches to directly target health-related applications. In particular, we go beyond the enzyme and pathway characterization in the approach design, pursuing applications of these unique enzymes in terms of flexibility to have a positive impact on human well-being. The proposed research is thus significant due to both implications for the field of natural product enzymology and utilities in biomedical and biotechnological research at the basic and translational levels.

总结 天然产物是研究、治疗甚至引发人类疾病的重要小分子， 它们通常具有对其生物活性至关重要的独特官能团。通过利用 通过合成这些功能的生物合成机制，可以增强、改变或减少 母体化合物的生物活性，并将生物合成机制应用于新的功能系统 组安装。为了实现这一目标，天然产物的化学逻辑和酶机制 生物合成需要充分表征和理解。本项目将侧重于功能特性 和机械审问的酶负责生物合成的不寻常的药效团的天然 产品包括末端炔、异腈和N-羟基三氮烯。独特的金属酶已经被 确定了重要的生物合成，包括膜结合的二铁依赖性双功能 用于炔生成的去饱和酶/乙炔酶，非血红素铁和α-酮戊二酸依赖性 用于异腈合成的氧化酶/脱羧酶，和由C-末端甲硫氨酰- tRNA合成酶样结构域和用于N-羟基三氮烯形成的N-末端cupin结构域。生化 对这些新发现的酶的机制和理解仍然很差， 在这个项目中。除了作为生物活性天然产物的弹头，这些功能还具有 不同的化学和物理性质，并用于各种生物正交化学转化， 化学生物学应用因此，我们将进一步探索其生物合成机械的应用， 这些“可点击”的功能在各种生物分子上的需求。研究战略在以下方面具有创新性： 新功能的选择及其独特但知之甚少的生物合成机制， 多学科研究工具，以及直接针对健康相关应用的目标导向方法。 特别是，我们超越了酶和途径表征的方法设计，追求 这些独特的酶在灵活性方面的应用对人类福祉产生积极影响。的 因此，由于对天然产物酶学领域的影响， 生物医学和生物技术研究的基础和转化水平的实用程序。