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: 10793148
• 负责人: Yi Tang
• 金额: $ 17.18万
• 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/10793148
• 关键词: Anabolism; Area; Biological; Chemical Structure; Cyclization; Educational process of instructing; Enzymatic Biochemistry; Enzymes; Family; Gene Cluster; Genome; Grant; Human; Investigation; Knowledge; Link; Metabolism; Methods; Microbial Genome Sequencing; Mining; Molds; Natural Products; Natural Resistance; Nature; Organism; Pathway interactions; Proteins; Renaissance; Research; Research Activity; Resistance; Resistance development; Structure; Therapeutic; Variant; 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创造了 天然产物发现的复兴。这种及时的组合为寻找 显示新结构和生物活性的天然产物。尽管有这样的 潜力，但仍然很难 i) 直接从生物合成基因预测产品结构 集群（BGC）。这是因为我们对天然产物中涉及的酶的了解 生物合成仍然有限，特别是对于高度编程的酶而言 真核生物，例如丝状真菌； ii) 优先考虑能够带来新的 BGC： 自然化学结构。这主要是由于该领域的重点是经过充分研究的自然 产品系列和核心生物合成酶； iii) 将生物活性与 基因组挖掘工作中的 BGC。这代表了基因组挖掘与传统技术之间的差距 基于表型筛选的发现，其中天然产物分离以生物学为指导 活动。 MIRA 赠款将通过综合研究计划解决这些限制 专注于真菌天然产物的发现和生物合成研究。 MIRA 项目的第一个总体领域是获得对核心的基本了解 参与真菌天然产物生物合成的酶。特别是，我们将 重点了解真菌 PKS 和 NRPS 的迭代编程规则。其他 核心酶编程规则的各个方面，包括环化和非规范域，将 被调查。我们还将研究真菌异常的剪裁酶活性 生物合成途径，重点是 PLP 依赖性酶和氧化酶。一些 将进行化合物驱动的生物合成研究。第二个一般领域 MIRA 项目旨在开发和完善基因组挖掘工具。最重要的研究 该领域的活动基于我们最近开发的抗性基因引导的目标基因组 采矿，其中我们使用天然产物目标的共簇、抗性变体 BGC 作为发现具有所需生物活性的天然产物的指南。这个策略还可以 用于将生物活性分配给已知的天然产物。这里的目标有两个—— 折叠：1）扩大可通过抗性基因识别的靶标列表，包括酶和 中心法则中的蛋白质、蛋白质运输、代谢等。这里我们将进行基因组分析 采矿和/或天然产物生物活性表征，将代谢物与目标联系起来； 2) 至 了解耐药机制，这将告诉我们大自然如何进化出耐药性 酶，并加深我们对如何克服潜在耐药性的理解。