PROTEIN AND METABOLIC ENGINEERING FOR BIOSYNTHESIS OF PHOSPHONIC ACID ANTIBIOTICS

膦酸抗生素生物合成的蛋白质和代谢工程

• 批准号: 7249593
• 负责人: Huimin Zhao
• 金额: $ 40.41万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7249593
• 关键词: Acetyl Coenzyme A; Address; Anabolism; Antibiotics; Antimalarials; Biological; Biological Factors; Biological Models; Citric Acid Cycle; Cloning; Engineering; Enzymes; Equilibrium; Escherichia coli; Exhibits; Fosfomycin; Gene Expression; Genes; Genetic Screening; Genomics; Glycolysis Pathway; Goals; Malaria; Medical; Metabolic; Methods; Modeling; Molecular Biology; Orthologous Gene; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Phosphonic Acids; Plants; Price; Production; Protein Overexpression; Proteins; Purpose; Rate; Research Personnel; Resistance; Resources; Roentgen Rays; Source; Streptomyces lividans; Structure; Treatment Efficacy; base; chemotherapeutic agent; combinatorial; combinatorial chemistry; directed evolution; engineering design; expression cloning; gene synthesis; human disease; improved; interest; microbial; microorganism; novel; phosphinothricin; programs; tool

The broad, long term objective of the proposed studies is to design and engineer phosphonic acid biosynthesis with the ultimate aim of developing a new framework for economical production of a wide variety of natural and unnatural natural products in Escherichia coli. Secondary metabolites - natural products - produced by plants and microorganisms are a prolific source for pharmaceutical drugs. Recent advances in molecular biology and genomics have revolutionized our ability to discover the biosynthetic pathways that synthesize natural products. However, since plants and microorganisms that produce interesting natural products are often poorly characterized or even uncultivable, the ability to manipulate their biosynthetic pathways is rather limited. In addition, natural products have evolved for the purpose other than treatment of human diseases. Although many natural products exhibit a diverse array of potent biological activities, their therapeutic efficacy often needs to be improved by further diversifying the core structures. More importantly, many of the natural product based drugs are produced in minute amounts in their native hosts, making the drugs very expensive. To address these issues, this proposal will use two important phosphonic acid (fosfomycin and FR-900098) biosynthetic pathways as model systemsto develop new methods and tools for economical biosynthesis of phosphonic acid antibiotics in E. coli. Such tools and methods should be generally applicable to the economical biosynthesis of a wide variety of natural products for biomedical applications. Moreover, microbial overproduction of FR900098, a promising antimalarial drug, may provide sufficient amounts at a low price for use in malaria-afflicted regions in the world. Finally, as a synergistic component of the program project, all relevant enzymes from these two pathways as well as other phosphonic acid (phosphinothricin, A53868, plumbemycin, rhizocticin, and SF2312) pathways will be overexpressed and purified for X-ray structural analysis by the Nair group. Some of them will also be biochemically and mechanistically characterized by the Metcalf, van der Donk and Kelleher groups.

拟议研究的广泛、长期目标是设计和工程膦酸 生物合成的最终目的是开发一个新的框架来经济生产广泛的 大肠杆菌中的多种天然和非天然天然产物。次生代谢物 - 天然 产品 - 由植物和微生物产生的产品是药品的丰富来源。最近的 分子生物学和基因组学的进步彻底改变了我们发现生物合成的能力 合成天然产物的途径。然而，由于产生 有趣的天然产物通常特征很差，甚至无法培养，操纵它们的能力 生物合成途径相当有限。此外，天然产物的进化目的除了 治疗人类疾病。尽管许多天然产物表现出多种有效的生物活性 活性，其治疗功效往往需要通过进一步多样化核心结构来提高。 更重要的是，许多基于天然产物的药物是在其天然产物中少量生产的。 宿主，使得药物非常昂贵。为了解决这些问题，本提案将使用两个重要的 膦酸（磷霉素和 FR-900098）生物合成途径作为模型系统来开发新的 在大肠杆菌中经济地生物合成膦酸抗生素的方法和工具。此类工具和 方法应普遍适用于多种天然产物的经济生物合成 用于生物医学应用。此外，微生物过量生产 FR900098（一种有前途的抗疟药）， 可以以低廉的价格提供足够的数量，供世界上受疟疾影响的地区使用。最后，作为一个 该计划项目的协同组成部分，来自这两个途径的所有相关酶以及 其他膦酸（草胺膦、A53868、铅霉素、根菌素和 SF2312）途径将 Nair 小组过表达并纯化用于 X 射线结构分析。其中一些也将是 Metcalf、van der Donk 和 Kelleher 小组在生物化学和机制上进行了表征。