Heteroatoms in natural products: Enzymes, pathways, and chemistry

前言：从生物体中分离出来的分子是化学上最复杂的化合物之一。我的项目旨在了解自然是如何构造天然产品的，我们使用化学和结构生物学作为主要工具。我们专注于如何制作特定的化学亚基序，这在发现生物催化剂和有针对性地分离特定类型的天然产品方面有应用。在这个NSERC探索计划中，我们关注的是大自然如何形成氮-氮(N-N)键。计划概述：这个计划建立在我们发现的第一个形成N-N键的生物合成酶的基础上。我们现在想要了解N-N键形成的详细机制，阐明植物中N-N键的形成途径，并发现新的含N-N键的天然产物。我们感兴趣的是酶N-N键形成的基本方面，以及我们的工作在生物催化、植物工程和靶向分子发现中的应用。第一部分：酶我们将测定N-N-键形成酶的晶体结构。通过结合体外研究、结构工作、机制研究和新变体的产生，我们预计将逐步揭开N-N键形成的机制。第二部分：途径我们将阐明N-N-键分子的途径。具体地说，我们将研究在黄瓜和亚麻中的β-吡唑-1-丙氨酸的生物合成。通过体内和体外研究，我们将发现新的酶，并研究它们如何产生这些独特的非蛋白生成氨基酸结构。第3部分：分子我们将使用一种有针对性的15N-核磁共振方法来筛选预计将产生哌嗪酸和N-亚硝基分子的细菌的部分，以发现新的天然产物，并为探索它们的生物学特性奠定基础。影响：我们的研究计划将探索生物合成的前沿，包括结构生物学、途径阐明和化合物发现。这些研究在以下方面有应用：1)开发用于化学合成的生物催化剂，因为酶正迅速取代传统的有机化学，用于精细化工生产；2)工程作物，使我们与生产不健康的非蛋白生成氨基酸有关的基因失活；3)有针对性地发现新的生物活性天然产品。HQP和EDI：我的综合培训计划将在未来5年内培训5名博士生、2名PDF和10名本科生。我致力于建立一个多元化的研究团队，其招聘策略将带来不同的声音。我通过实施日常行动来努力实现包容性，以确保所有团队成员的意见得到倾听和包括在内。最后，我倡导平等的机会，以确保每个人的个人和职业发展。我的项目为学生提供化学、分子生物学和结晶学方面的培训，并鼓励学生的创造力和独立性的发展，所有这些都将使学生在不同的职业领域取得成功，如学术界、生物技术。

Introduction: Molecules isolated from living organisms are among the most chemically complex compounds. My program aims to understand how nature constructs natural products, and we use chemistry and structural biology as major tools. We focus on how specific chemical sub-motifs are made, which has applications in the discovery of biocatalysts and targeted isolation of specific types of natural products. In this NSERC Discovery program, we focus on how nature forms nitrogen-nitrogen (N-N) bonds. Program Overview: This program builds on our discovery of the first biosynthetic enzyme to form an N-N bond. We now want to understand the detailed mechanisms of N-N bond formation, elucidate pathways to N-N bonds in plants, and discover new N-N-bond-containing natural products. We are interested in both fundamental aspects of enzymatic N-N bond formation, and applications of our work in biocatalysis, plant engineering, and targeted molecule discovery. Part 1: Enzymes We will determine the crystal structures of N-N-bond-forming enzymes. By combining in vitro studies, structural work, mechanistic studies, and generation of new variants, we expect to unravel the step-by-step mechanisms of N-N bond formation. Part 2: Pathways We will elucidate pathways to N-N-bond-containing molecules. Specifically, we will study the biosynthesis of ß-pyrazol-1-ylalanine in cucumber and linatine in flax. Through in vivo and in vitro studies, we will find new enzymes and study how they generate these unique non-proteinogenic amino acid structures. Part 3: Molecules We will employ a targeted, 15N-NMR-approach to screen fractions from bacteria predicted to produce piperazic acid and N-nitroso-containing molecules to discover new natural products and set the stage for exploration of their biological properties. Impact: Our research program will explore the frontiers of biosynthesis, including structural biology, pathway elucidation, and compound discovery. These studies have application in 1) development of biocatalysts for chemical synthesis, as enzymes are rapidly replacing traditional organic chemistry for fine chemical production; 2) engineering of crop plants to inactivate genes we link to production of unhealthy non-proteinogenic amino acids; 3) targeted discovery of new bioactive natural products. HQP and EDI: My comprehensive training plan will train 5 PhD students, 2 PDFs, and 10 undergraduate students over the next 5 years. I work to build a diverse research team with a recruitment strategy that brings diverse voices. I work toward inclusivity by implementing day-to-day actions to ensure all team members are heard and included. Finally, I champion equal opportunity to ensure personal and professional growth for everyone. My program gives students training in chemistry, molecular biology, and crystallography, and encourages creativity and development of independence, all of which will position students for success in diverse careers e.g. academia, biotechnology.