MIRA: Enzymology and Self-Resistance of Natural Product Biosynthesis

MIRA：天然产物生物合成的酶学和自身抗性

• 批准号: 10163012
• 负责人: Yi Tang
• 金额: $ 58.61万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10163012
• 关键词: Address; Anabolism; Area; Biological; Chemical Structure; Cyclization; Enzymatic Biochemistry; Enzymes; Family; Gene Cluster; Genome; Grant; Human; Investigation; Knowledge; Lead; Link; Metabolism; Methods; Microbial Genome Sequencing; Mining; Molds; Natural Products; Natural Resistance; Nature; Organism; Pathway interactions; Phenotype; Proteins; Renaissance; Research; Research Activity; Resistance; Resistance development; Structure; Therapeutic; Time; Variant; base; bioactive natural products; enzyme activity; microbial genome; programs; protein transport; resistance gene; resistance mechanism; synthetic biology; tool

ABSTRACT Recent progresses in microbial genome sequencing and synthetic biology have created a renaissance in natural product discovery. This timely combination offers great promise to find natural products displaying new structures and biological activities. Notwithstanding such potential, it remains difficult to i) predict product structures directly from biosynthetic gene clusters (BGCs). This is because our knowledge of enzymes that are involved in natural product biosynthesis remains limited, especially with regard to the highly programmed enzymes from eukaryotic organisms such as filamentous fungi; ii) prioritize BGCs that can lead to new-to- nature chemical structures. This is primarily due to the focus of the field on well-studied natural product families and core biosynthetic enzymes; and iii) connect the biological activity with BGCs in genome mining efforts. This represents a gap between genome mining and traditional phenotypical screen-based discovery in which natural product isolation is guided by biological activity. This MIRA grant will address these limitations with a comprehensive research program focused on fungal natural product discovery and biosynthetic investigation. The first general area of this MIRA project is to gain fundamental understanding of core enzymes that participate in the biosynthesis of fungal natural products. In particular, we will focus on understanding the iterative programming rules of fungal PKSs and NRPSs. Other aspects of core enzyme programming rules, including cyclization and noncanonical domains will be investigated. We will also investigate the unusual tailoring enzyme activities of fungal biosynthetic pathways, with emphasis on PLP-dependent and oxidative enzymes. A number of compound driven biosynthetic investigations will be conducted. The second general area of this MIRA project is to develop and refine tools for genome mining. The most important research activity in this area is based on our recently developed resistance gene guided target genome mining, in which we use an co-clustered, resistant variant of the natural product target in the BGC as a guide to discover natural product of desired biological activity. This strategy can also be used to assign biological activities to known natural products. The objectives here are two- fold: 1) to expand the list of targets that may be identified via resistance gene, to enzymes and proteins in the central dogma, protein transport, metabolism, etc. Here we will perform genome mining and/or natural product bioactivity characterization to link metabolites to targets; and 2) to understand the mechanism of resistance, which will teach us how Nature evolves resistant enzymes, and refine our understanding of how to overcome potential resistance.

抽象的 微生物基因组测序和合成生物学的最新进展已创造 自然产品发现的文艺复兴。这种及时的组合提供了很大的希望 展示新结构和生物活动的天然产品。尽管如此 潜力，很难i）直接从生物合成基因预测产品结构 集群（BGC）。这是因为我们对涉及天然产品的酶的了解 生物合成仍然有限，特别是关于来自 真核生物，例如丝状真菌； ii）优先考虑BGC，这可能导致新到新的 自然化学结构。这主要是由于该领域的重点放在研究良好的自然上 产品家族和核心生物合成酶； iii）将生物活性与 BGC在基因组采矿工作中。这代表了基因组开采与传统之间的差距 基于表型筛查的发现，其中自然产物隔离由生物学指导 活动。 MIRA赠款将通过一项全面的研究计划解决这些限制 专注于真菌天然产品发现和生物合成研究。 这个MIRA项目的第一个一般领域是获得对核心的基本理解 参与真菌天然产品的生物合成的酶。特别是，我们会 专注于理解真菌PKS和NRPS的迭代编程规则。其他 核心酶编程规则的各个方面，包括环化和非规范领域 被调查。我们还将研究真菌的异常剪裁酶活性 生物合成途径，重点是依赖PLP的酶和氧化酶。许多 将进行复合驱动的生物合成研究。第二个一般区域 MIRA项目将开发和完善基因组采矿的工具。最重要的研究 该领域的活性基于我们最近开发的抗性基因引导的靶基因组 采矿，在其中使用自然产品目标的共聚类，抗性变体 BGC作为发现所需生物活性的天然产物的指南。这个策略也可以 用于将生物活性分配给已知的天然产品。这里的目标是两个 折：1）扩大可以通过电阻基因，酶和酶鉴定的靶标列表 中央教条中的蛋白质，蛋白质传输，代谢等。我们将执行基因组 采矿和/或天然产物生物活性表征将代谢产物与靶标联系起来；和2）到 了解抵抗的机制，这将教会我们自然如何发展抵抗力 酶，并完善我们对如何克服潜在抗性的理解。