PROTEIN AND METABOLIC ENGINEERING FOR BIOSYNTHESIS OF PHOSPHONIC ACID ANTIBIOTICS

拟议研究的广泛、长期目标是设计和设计磷酸 生物合成，最终目的是开发一种新的经济生产框架 大肠杆菌中天然和非天然产物的种类。次生代谢物--天然 产品-由植物和微生物生产的产品是制药药物的丰富来源。近期 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，由于产生的植物和微生物 有趣的天然产品往往特性不佳，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，天然产品进化的目的不是为了 人类疾病的治疗。尽管许多天然产品展示了一系列不同的有效生物 为了更好地开展活动，它们的治疗效果往往需要通过进一步使核心结构多样化来提高。 更重要的是，许多以天然产物为基础的药物都是在其本土生产的微量药物 主办方，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 磷酸(磷霉素和FR-900098)生物合成途径作为开发新系统的模式系统 在大肠杆菌中经济地生物合成膦酸类抗生素的方法和工具。这些工具和 方法应普遍适用于各种天然产品的经济生物合成 用于生物医学应用。此外，前景看好的抗疟疾药物FR900098的微生物生产过剩， 可提供足够数量的低价格供世界疟疾肆虐地区使用。最后，作为一个 计划项目的协同部分，来自这两个途径的所有相关酶以及 其他磷酸(膦硫蛋白、A53868、白霉素、根霉素和SF2312)途径将是 过表达和纯化，用于Nair小组的X-射线结构分析。其中一些人也将成为 Metcalf、van der Donk和Kelleher基团具有生物化学和机械特性。

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.