Heteroatoms in natural products: Enzymes, pathways, and chemistry

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。我的计划旨在了解自然如何构建天然产物，我们使用化学和结构生物学作为主要工具。我们专注于特定的化学亚基序是如何制作的，这在生物催化剂的发现和特定类型的天然产物的靶向分离中有应用。在这个NSERC发现计划中，我们专注于自然界如何形成氮-氮（N-N）键。计划概述：该计划建立在我们发现第一种形成N-N键的生物合成酶的基础上。我们现在希望了解N-N键形成的详细机制，阐明植物中N-N键的途径，并发现新的含N-N键的天然产物。我们对酶促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都很感兴趣。第一部分：酶我们将确定N-N键形成酶的晶体结构。通过结合体外研究，结构工作，机理研究和新变体的产生，我们期望解开N-N键形成的逐步机制。第2部分：途径我们将阐明含N-N键分子的途径。具体而言，我们将研究黄瓜中的β-吡唑-1-基丙氨酸和亚麻中的亚麻碱的生物合成。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质氨基酸结构。第三部分：分子我们将采用有针对性的15 N-NMR方法，从预测产生哌嗪酸和含N-亚硝基分子的细菌中筛选馏分，以发现新的天然产物，并为探索其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。这些研究可应用于1）开发用于化学合成的生物催化剂，因为酶正在迅速取代传统的有机化学用于精细化学品生产; 2）工程化作物植物以抑制我们与不健康的非蛋白质氨基酸生产相关的基因; 3）有针对性地发现新的生物活性天然产物。HQP和EDI：我的综合培训计划将在未来5年内培养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，其招聘策略可以带来不同的声音。我通过实施日常行动来实现包容性，以确保所有团队成员都能听到并被包括在内。最后，我倡导平等机会，以确保每个人的个人和职业成长。我的课程为学生提供化学，分子生物学和晶体学方面的培训，并鼓励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生涯中取得成功，例如学术界，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.