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：
10830540
负责人：
Bo Li
金额：
$ 0.73万
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

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，可以重新使用以高效且可持续的综合工业和药物化合物。识别新型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的抗生素荧光素C，由铜绿假单胞菌，这是多药耐药性医院感染的主要原因。经过研究铜绿假单胞菌如何制作荧光素C，我们确定了一种不寻常的生物合成途径涉及新型酶化学。例如，新的血红素的两个铁依赖性酶氧合酶样酶家族催化碳切除和氧化脱羧和 N-羟基化。腺苷糖酸酯裂解酶家族的两个裂解酶表现出分别建立和破坏碳硫键的新活动。我们的化学条件确定的已经扩大了这些酶家族的催化能力。我们假设这一点荧光素C生物合成酶使用新型的催化机制，可以重新使用或设计为生成新分子。为了检验这一假设，我们将使用酶动力学，结构生物学，光谱和合成化学工具，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推进对各自的酶家族如何催化多样化的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的使用。