Elucidate Mechanisms of Quinolone Alkaloid Biosynthesis via Iron(II)/2-Oxoglutarate Dependent Enzymes: Diverse, but Controlled Reactivity

通过铁 (II)/2-氧戊二酸依赖性酶阐明喹诺酮生物碱生物合成的机制：多样但受控的反应性

• 批准号: 9753300
• 负责人: Yisong Guo
• 金额: $ 31.05万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753300
• 关键词: Alkaloids; Anabolism; Anti-Bacterial Agents; Antiviral Agents; Area; Aspergillus nidulans; Biochemical; Biological; Biological Assay; Biomedical Engineering; Catalysis; Cations; Chemicals; Comparative Study; Crystallography; Cytochrome P450; Deuterium; Development; Enzymes; Epigenetic Process; Exhibits; Family; Gene Expression Regulation; Goals; HIV; Hydrogen; Hydroxylation; Intercept; Iron; Kinetics; Knowledge; Label; Lead; Libraries; Logic; Malignant Neoplasms; Metabolism; Methods; Molecular; Molecular Cloning; Mononuclear; Natural Products; Nature; Organic Synthesis; Outcome; Oxidants; Oxygen; Pathway interactions; Pharmacologic Substance; Play; Preparation; Proteins; Quinolones; Reaction; Regulation; Research; Role; Scheme; Site; Solvents; Structure; Structure-Activity Relationship; Techniques; Time; Variant; Viral Cancer; X-Ray Crystallography; alpha ketoglutarate; analog; anti-cancer; anticancer activity; antimicrobial; base; design; drug development; drug discovery; electronic structure; functional group; improved; insight; interest; member; non-Native; novel; scaffold; simulation

Project Summary/Abstract 2-Oxoglutarate (2OG) dependent nonheme mononuclear iron (NHM-Fe) enzymes catalyze an exceedingly broad scope of reactions that are involved in key chemical transformations of many important biological pathways, such as gene regulation, epigenetics, and natural product biosynthesis. Although detailed mechanistic understandings of the canonical hydroxylation reactivity found in 2OG/NHM-Fe enzymes have been developed in recent years, it remains unknown how this hydroxylation paradigm can fully explain non- hydroxylation reactivity in this family of enzymes, such as desaturation and epoxidation. Furthermore, given the catalytic abilities of 2OG/NHM-Fe enzymes to construct pharmaceutically valuable molecular scaffolds, exploiting these enzymes for biocatalysis applications represents an attractive but under developed area for expanding natural product based compound libraries. In this proposal, we seek to provide critical improvements on these under developed areas through the studies of AsqJ, a novel multifunctional 2OG/NHM- Fe enzyme that is involved in Viridicatin-type quinolone alkaloid biosynthesis in Aspergillus nidulans. AsqJ catalyzes a chemically interesting sequential desaturation/epoxidation reaction to construct Viridicatin core structure, which represents a chemically unexplored strategy for Viridicatin synthesis. A multi-faceted experimental method will be utilized to elucidate AsqJ reaction mechanisms, which consists of organic synthesis, molecular cloning, biochemical assays, protein crystallography, pre-steady state kinetics, and advanced spectroscopic techniques. This method will be further supplemented with molecular dynamic simulations to generate molecular level understandings of the AsqJ catalysis. It is expected that the proposed research will provide critical improvements to the mechanistic understandings of desaturation and epoxidation, two chemically challenging but under explored reactions catalyzed by 2OG/NHM-Fe enzymes, and further explore mechanism based bioengineering approach to access viridicatin-type scaffolds.

项目总结/文摘