Mechanisms of nonribosomal peptide natural product biosynthesis

非核糖体肽天然产物生物合成机制

• 批准号: 7802060
• 负责人: Steven D Bruner
• 金额: $ 27.09万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-10 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7802060
• 关键词: Address; Amino Acids; Amino Acids Activation; Ammonia-Lyases; Anabolism; Antibiotics; Architecture; Biochemical; Biological Factors; Catalysis; Chemicals; Chemistry; Coenzyme A; Complex; Crystallization; Dioxygen; Enediyne Antitumor Antibiotic; Engineering; Enzymatic Biochemistry; Enzymes; Exhibits; Family; Generations; Goals; Ligase; Metabolic Pathway; Metals; Methodology; Microbe; Movement; Nucleic Acid-Independent Peptide Biosynthesis; Organic Synthesis; Organism; Oxygen; Oxygenases; Pathway interactions; Peptides; Pharmaceutical Preparations; Plants; Process; Property; Prosthesis; Research; Route; Source; Specificity; Staging; Structure; System; Techniques; Teicoplanin; Therapeutic; Tyrosine; Vancomycin; X-Ray Crystallography; analog; base; catalyst; chemical binding; cofactor; design; fascinate; in vivo; insight; interest; leinamycin; member; novel; novel therapeutics; peptide synthase; programs; protein protein interaction; public health relevance; tool

DESCRIPTION (provided by applicant): Peptide natural products represent a diverse class of important therapeutics. The biosynthetic enzymes responsible for constructing nonribosomal peptides have complex structural architecture and frequently carry out difficult chemical transformations. Manipulation of biosynthetic pathways through in vivo engineering or chemoenzymatic techniques is a promising approach to the generation of novel therapeutics. Presented is our program to elucidate the mechanisms and provide structural information for key steps in the biosynthesis of nonribosomal peptides. The project will examine two important aspects of the biosynthetic methodology: the biosynthesis of nonproteinogenic amino acid building blocks and the selection and loading of amino acids onto the synthetase machinery. We will apply an interdisciplinary combination of X-ray crystallography, organic synthesis and mechanistic enzymology. The results will be applied to the rational engineering of novel amino acid building blocks and is part of our long term goal of understanding the complex mechanisms natural product assembly-line biosynthesis. Systems under study are selected for both exhibited novel enzymology and importance in the biosynthesis of therapeutically important molecules. The proposal describes three enzyme systems, each contributing unique information toward the overall project goals. 1) Nonproteinogenic amino acids are key components of the vancomycin class of antibiotics. An important step in the biosynthesis of 3,5-dihydroxyphenylglycine is the dioxygenation catalyzed by the enzyme DpgC. DpgC is a member of a very small group of cofactor/metal independent oxygenases and has unique chemistry and structure. 2) ?-Amino acids are important building blocks in a wide range of natural and synthetic compounds, including the antitumor/antibiotic enediynes. A novel aminomutase, SgcC4, containing the rare cofactor 4-methylideneimidazolone catalyzes the 1,2-amino shift of ?-tyrosine to generate ?-tyrosine. 3) The structural basis of domain/domain interactions of nonribosomal peptide machinery will be examined using designed synthetic analogs as tethering agents. We will apply this approach to the stand-alone didomain constructs responsible for activation and loading of amino acids. PUBLIC HEALTH RELEVANCE: A large percentage of useful therapeutics are derived from natural sources like microbes and plants. Our research program is interested in understanding the details of how simple organisms developed complex chemistry to produce important drug molecules. This effort will assist in efforts to manipulate natural product producers as a route to the discovery of novel drugs with desired properties.

描述（由申请人提供）：肽天然产物代表了多种重要的治疗方法。负责构建非核糖体肽的生物合成酶具有复杂的结构，并且经常进行困难的化学转化。通过体内工程或化学酶技术操纵生物合成途径是产生新疗法的一种有前途的方法。提出的是我们的计划，旨在阐明非核糖体肽生物合成关键步骤的机制并提供结构信息。该项目将研究生物合成方法的两个重要方面：非蛋白氨基酸结构单元的生物合成以及氨基酸的选择和加载到合成酶机器上。我们将应用 X 射线晶体学、有机合成和机械酶学的跨学科组合。研究结果将应用于新型氨基酸结构单元的合理工程，并且是我们了解天然产物装配线生物合成复杂机制的长期目标的一部分。选择正在研究的系统是因为其在治疗重要分子的生物合成中表现出新颖的酶学和重要性。该提案描述了三个酶系统，每个系统都为总体项目目标提供独特的信息。 1) 非蛋白氨基酸是万古霉素类抗生素的关键成分。 3,5-二羟基苯基甘氨酸生物合成的一个重要步骤是 DpgC 酶催化的双氧化作用。 DpgC 是一小群不依赖于辅因子/金属的加氧酶的成员，具有独特的化学和结构。 2) α-氨基酸是多种天然和合成化合物的重要组成部分，包括抗肿瘤/抗生素烯二炔。一种新型氨基变位酶 SgcC4，含有稀有辅因子 4-亚甲基咪唑酮，可催化 β-酪氨酸的 1,2-氨基转移生成 β-酪氨酸。 3) 将使用设计的合成类似物作为束缚剂来检查非核糖体肽机器的结构域/结构域相互作用的结构基础。我们将这种方法应用于负责氨基酸激活和加载的独立双结构域构建体。公共卫生相关性：很大一部分有用的治疗方法源自微生物和植物等天然来源。我们的研究项目有兴趣了解简单生物体如何发展复杂化学来生产重要药物分子的细节。这项工作将有助于操纵天然产物生产商，作为发现具有所需特性的新药物的途径。