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

基本信息

批准号：
10653015
负责人：
Wenjun Zhang
金额：
$ 33.24万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
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/10653015
关键词：
Acids Acyl Carrier Protein Alkynes Anabolism Binding Biochemical Biological Biological Models Biology Biotechnology C-terminal Carboxy-Lyases Chemicals Chemistry Engineering Enzymatic Biochemistry Enzymes Family Gene Cluster Generations Genome Goals Health Human Interdisciplinary Study Iron Isotope Labeling Knowledge Libraries Logic Membrane Methionine-tRNA Ligase Mining Mycobacterium tuberculosis N Domain N-terminal Natural Products Nature Nitrogen Nitrous Acid Organic Synthesis Oxidases Parents Pathway interactions Personal Satisfaction Pharmacologic Substance Prevalence Public Health Reaction Research Streptomyces aureofaciens Structure Substrate Specificity System Testing Virulence Factors Work alpha ketoglutarate bioactive natural products chemical property desaturase design experimental study feeding flexibility functional group human disease improved in vivo innovation metalloenzyme novel pharmacophore physical property polyketide synthase polyketides scaffold small molecule synthetic drug tool

项目摘要

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.

天然产物是研究、治疗甚至引起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，通过利用合成这些功能的生物合成机制，可以增强、为了实现这一目标，需要充分表征和理解天然产物生物合成的化学逻辑和酶机制。负责天然产物不常见药效基团（包括末端炔、异腈和 N-羟基三氮烯）生物合成的酶的功能表征和机械研究已鉴定出对其生物合成非常重要的独特金属酶，包括膜结合的双铁依赖性双功能去饱和酶/乙酰化酶。对于炔烃的生成，非血红素铁和 α-酮戊二酸依赖性氧化酶/脱羧酶用于异腈合成的双结构域酶，由 C 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 端 cupin 结构域组成。 -该项目的深入研究除了作为生物活性天然产物的弹头外，这些功能还具有独特的化学和物理性质，并可用于各种生物正交化学转化。因此，我们将进一步探索其生物合成机制的应用，以根据需要在各种生物分子上安装这些“可点击”功能。该研究策略在新颖功能的选择及其独特但知之甚少的生物合成机制方面具有创新性。特别是，我们在方法设计中超越了酶和途径表征，追求这些独特酶的灵活性应用，以产生积极影响。对人类因此，由于对天然产物酶学领域以及基础和转化层面的生物医学和生物技术研究的实用性的影响，拟议的研究具有重要意义。