Excellence in Research: Biosynthetic investigation of manzamine class alkaloids

卓越研究：曼扎明类生物碱的生物合成研究

• 批准号: 2302454
• 负责人: Madan Kharel
• 金额: $ 46.03万
• 依托单位: University of Maryland Eastern Shore
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2302454&HistoricalAwards=false
• 关键词: Excellence; Research; Biosynthetic; investigation; manzamine

Manzamines are members of marine alkaloid natural products. They possess complex structures and exhibit a number of biological activities, including anticancer and antibacterial activities. Lack of knowledge of the molecular and biochemical basis for the production of these alkaloids has been a major barrier to the bioengineering of new alkaloid derivatives. This project will provide such knowledge and will serve as the foundation for bioengineering of new manzamine derivatives with potential applications in medicine, agriculture and biotechnology. This project will also provide extensive research experiences for University of Maryland Eastern Shore undergraduate and graduate students, many of whom are from groups underrepresented in STEM, and thus the work will contribute to development of a diverse STEM workforce. Manzamines represent a structurally distinct group of marine alkaloid natural products with little resemblance to other alkaloids. The structural uniqueness of manzamines, including manzamine A arises from the assembly of a complex pentacyclic core attached to a beta-carboline unit. Although over a hundred manzamines that exhibit a wide range of biological activities have been isolated primarily from marine sponges, how organisms assemble building blocks and intermediates to furnish complex structural scaffolds of manzamines has remained a mystery. The main objective of the proposed project is to identify biosynthetic building blocks and pathway intermediates and elucidate key biosynthetic steps involved in the production of manzamine A. The proposed project utilizes a multi-pronged experimental approach including: (A) isotope incorporation experiments to identify biosynthetic building blocks of the beta-carboline and the pentacyclic ring, (B) experiments to identify the enzyme involved in the attachment of these two structural units, (C) complete genome sequence analysis of Micromonospora sp. M42 for identifying manzamine biosynthetic locus, and (D) heterologous production and characterization of select enzymes involved in the manzamine A biosynthetic pathway. The results garnered through this project will pave the way for the discovery of other manzamines through bioinformatic analysis. Manzamine A biosynthetic enzymes with potentially new catalytic activities will be an invaluable addition to the existing repertoire of tools for the further advancement of research projects concerning biosynthetic pathway engineering/synthetic biology, particularly for the production of new manzamine analogs. The proposed project thus offers a biological alternative to the synthetic production of new manzamine alkaloids.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

Manzamines是海洋生物碱天然产物的成员。它们具有复杂的结构，并表现出许多生物活性，包括抗癌和抗菌活性。缺乏知识的分子和生物化学基础的生产这些生物碱已成为一个主要的障碍，生物工程的新生物碱衍生物。该项目将提供这些知识，并将作为新的manzamine衍生物的生物工程的基础，在医学，农业和生物技术方面具有潜在的应用。该项目还将为马里兰州东海岸大学的本科生和研究生提供广泛的研究经验，其中许多人来自STEM代表性不足的群体，因此这项工作将有助于发展多元化的STEM劳动力。Manzamines代表了一组结构独特的海洋生物碱天然产物，与其他生物碱几乎没有相似之处。曼扎胺（包括曼扎胺A）的结构独特性源于与β-咔啉单元连接的复杂五环核的组装。虽然已经从海绵中分离出了100多种具有广泛生物活性的manzamines，但生物体如何组装积木和中间体以提供manzamines的复杂结构支架仍然是一个谜。该项目的主要目标是确定生物合成的基本单元和途径中间体，并阐明生产曼扎胺A的关键生物合成步骤。拟议项目采用多管齐下的实验方法，包括：（A）同位素掺入实验，以鉴定β-咔啉和五环的生物合成构件，（B）实验，以鉴定参与这两个结构单元连接的酶，（C）小单孢菌属M42的完整基因组序列分析，以鉴定曼扎胺生物合成基因座，和（D）异源生产和表征参与manzamine A生物合成途径的选择酶。通过该项目获得的结果将为通过生物信息学分析发现其他manzamines铺平道路。具有潜在的新催化活性的曼扎明A生物合成酶将是对现有工具库的宝贵补充，用于进一步推进关于生物合成途径工程/合成生物学的研究项目，特别是用于生产新的曼扎明类似物。因此，拟议的项目提供了一个生物替代新的manzamine生物碱的合成生产。这个奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。