Mechanisms of unusual enzymes in the biosynthesis of a copper-containing antibiotic

含铜抗生素生物合成中异常酶的机制

基本信息

批准号：
10567957
负责人：
Bo Li
金额：
$ 30.67万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-20 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10567957
关键词：
Amines Anabolism Anaerobic Bacteria Antibiotics Binding Biomedical Engineering Carbon Catalysis Chemical Structure Chemistry Collaborations Copper Crystallization Cysteine Data Decarboxylation Dioxygenases Electron Spin Resonance Spectroscopy Engineering Enzyme Kinetics Enzymes Equilibrium Excision Exhibits Family Fumarates Hydrogen Industrialization Infection Investigation Ions Iron Lifting Ligand Binding Ligands Lyase Medicine Metal Ion Binding Metals Microbe Monitor Multi-Drug Resistance N hydroxylation Natural Products Nature Nitric Oxide Nosocomial Infections Oximes Oxygen Pathway interactions Pharmacologic Substance Play Production Pseudomonas aeruginosa Reaction Reducing Agents Roentgen Rays Siderophores Spectrum Analysis Structure Sulfur Synthesis Chemistry Testing Variant Virulence Work absorption adenylosuccinate lyase analog base carboxylate college enzyme mechanism fluopsin human pathogen innovation insight microorganism novel pathogen pathogenic bacteria small molecule stoichiometry structural biology synthetic biology tool

项目摘要

Abstract Enzymes are powerful biocatalysts that may be repurposed for efficient and sustainable synthesis of industrial and pharmaceutical compounds. Identifying novel enzyme-catalyzed reactions and understanding their catalytic mechanisms is essential for expanding the biocatalytic toolbox. We have recently discovered a unique copper-containing antibiotic, fluopsin C, which is produced by Pseudomonas aeruginosa, a leading cause of multidrug-resistant nosocomial infections. By studying how P. aeruginosa makes fluopsin C, we identified an unusual biosynthetic pathway that involves novel enzyme chemistry. For instance, two iron-dependent enzymes of a new heme- oxygenase-like enzyme family catalyze a carbon excision and an oxidative decarboxylation and N-hydroxylation respectively. Two lyases of the adenylosuccinate lyase enzyme family exhibit novel activities of making and breaking carbon-sulfur bonds respectively. The chemistries we identified have expanded the catalytic capabilities of these enzyme families. We hypothesize that the fluopsin C biosynthetic enzymes use novel catalytic mechanisms and can be repurposed or engineered to generate new molecules. To test this hypothesis, we will use a combination of enzyme kinetics, structural biology, spectroscopy, and synthetic chemistry tools, with a focus on three enzymes in fluopsin C biosynthesis. Studies of these distinct enzymes will advance the fundamental understanding of how their respective enzyme family catalyzes diverse chemistry. Defining the mechanisms and substrate scope of the enzymes will allow them to be further explored for biocatalytic use.

抽象的酶是强大的生物催化剂，可以重新用于高效和可持续的合成工业和药物化合物。识别新的酶催化反应和了解它们的催化机制对于扩展生物催化工具箱至关重要。我们最近发现了一种独特的含铜抗生素，fluopsin C，它是由铜绿假单胞菌，多重耐药医院感染的主要原因。经过通过研究铜绿假单胞菌如何产生氟视蛋白 C，我们发现了一种不寻常的生物合成途径涉及新颖的酶化学。例如，一种新血红素的两种铁依赖性酶- 加氧酶样酶家族催化碳切除和氧化脱羧分别为N-羟基化。腺苷琥珀酸裂解酶家族的两种裂解酶表现出分别形成和破坏碳硫键的新活性。我们的化学已鉴定的酶扩大了这些酶家族的催化能力。我们假设氟视蛋白 C 生物合成酶采用新颖的催化机制，可以重新利用或被设计来产生新分子。为了检验这个假设，我们将使用以下组合酶动力学、结构生物学、光谱学和合成化学工具，重点是氟视素 C 生物合成中的三种酶。对这些不同酶的研究将推动对各自酶家族如何催化不同化学反应的基本了解。定义酶的机制和底物范围将使它们进一步探索用于生物催化用途。