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

• 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
• 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

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.

摘要 酶是强大的生物催化剂，可用于高效和可持续的合成 工业和医药化合物的生产。识别新的酶催化反应， 了解它们的催化机制对于扩大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，荧光素C，它是由 铜绿假单胞菌，多重耐药医院感染的主要原因。通过 通过研究铜绿假单胞菌如何制造荧光素C，我们发现了一种不寻常的生物合成途径， 涉及新的酶化学。例如，一种新血红素的两种铁依赖酶- 加氧酶样酶家族催化碳切除和氧化脱羧， N-羟基化。腺苷酸琥珀酸裂解酶家族的两种裂解酶表现出 分别具有生成和断裂碳-硫键的新活性。我们所使用的化学物质 已经确定的这些酶家族的催化能力扩大。我们假设 荧光素C生物合成酶使用新的催化机制， 被改造以产生新的分子为了验证这个假设，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 荧光素C生物合成中的三种酶。对这些不同酶的研究将促进 基本了解它们各自的酶家族如何催化不同的化学反应。 确定酶的机制和底物范围将使它们能够进一步 探索用于生物催化用途。