Discovery of catalytic tools for isoprenoid production

发现类异戊二烯生产的催化工具

基本信息

批准号：
RGPIN-2017-04874
负责人：
Ro, DaeKyun
金额：
$ 2.84万
依托单位：
University of Calgary
依托单位国家：
加拿大
项目类别：
Discovery Grants Program - Individual
财政年份：
2019
资助国家：
加拿大
起止时间：
2019-01-01 至 2020-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=687321
关键词：
Discovery catalytic tools isoprenoid production

项目摘要

The isoprenoids are a large and structurally diverse class of natural products with more than 55,000 known compounds. Many isoprenoids have been used as flavors, pharmaceuticals, and industrial polymers. All isoprenoids are currently supplied either by purification from plants or by chemical synthesis involving petrochemicals. However, the yield from natural sources is often inadequate to meet industrial demands, and chemical synthesis is not desirable due to environmental perturbation. To overcome these, the enzymatic production of isoprenoids using biological systems has become an attractive alternative to the traditional purification and chemical synthesis; however, our current knowledge of isoprenoid metabolism remains insufficient to realize the biological supply of isoprenoids. ******The long-term goal of my research program is to supply isoprenoid products by sustainable biological methods, thus replacing the petrochemical-derived products. To advance the knowledge of isoprenoid metabolism, I propose to study two subclasses of isoprenoid natural products - sesquiterpene lactones (SLs) and natural rubber (NR). SLs have been used as building blocks for many pharmaceuticals while NR is an indispensable biopolymer for hundreds of industrial products. Precise mechanisms of SL and NR biosynthesis remain elusive, and integrative approaches combining biochemistry, genetics, and genomics will be used to elucidate the mysteries of SL and NR biosynthesis. ******First, how stereoselective biosynthesis of various SLs is achieved in nature will be studied by isolating and characterizing enzymes from two medicinal plant species. Stereoselective synthesis of chemicals is essential in the pharmaceutical industry, and thus this work will provide biological methods to produce stereoselective chemicals. Second, combinatorial uses of plant enzymes will be attempted in microbes to produce novel SLs that are not present in nature, based on our discovery of one promiscuous enzyme in SL biosynthesis. More than 100 combinations of enzymes will be synthetically made in yeast, and the resulting new SL products will be identified and purified for structural studies. Ultimately, these new SLs will be examined for possible anti-bacterial activities. Third, as we do not fully understand how NR is biosynthesized in plants, in part due to gaps in our knowledge of several protein components in the NR pathway, additional necessary proteins for NR biosynthesis will be identified and characterized from plants using biochemical and genetic approaches. The newly identified protein component(s) can be used to devise alternative bio-systems that can produce NR outside of the tropical para-rubber trees, thereby securing polymer sustainability. Collectively, the proposed research will lay a foundation of knowledge for a sustainable supply of isoprenoid products critically needed in our society.**

类异on-是一种大型且结构上多样的天然产物，具有超过55,000种已知化合物。许多类异on-已用作口味，药物和工业聚合物。目前，所有类异丙生素都是通过植物纯化或涉及石化物质的化学合成来提供的。但是，自然来源的产量通常不足以满足工业需求，并且由于环境扰动而无法获得化学合成。为了克服这些，使用生物系统的类异丙定酶的酶促产生已成为传统纯化和化学合成的有吸引力的替代品。但是，我们目前对异丙代谢的知识仍然不足以实现类吸收的生物供应。 *****我的研究计划的长期目标是通过可持续的生物学方法提供类异丙裔产品，从而取代石化衍生的产品。为了促进异丙裔代谢的知识，我建议研究异丙酸酯天然产物的两个亚类-Estementementemententement -secentrenepene Lactones（SLS）和天然橡胶（NR）。 SLS已被用作许多药品的基础，而NR是数百种工业产品必不可少的生物聚合物。 SL和NR生物合成的精确机制仍然难以捉摸，将生物化学，遗传学和基因组学结合的综合方法将用于阐明SL和NR生物合成的奥秘。 ******首先，如何通过分离和表征从两种药用植物物种的酶来研究各种SLS的立体选择性生物合成。化学物质的立体选择性合成在制药行业至关重要，因此这项工作将提供生产立体选择化学物质的生物学方法。其次，基于我们在SL生物合成中发现一种混杂酶，将尝试在微生物中尝试在微生物中尝试产生不存在的新型SLS。将在酵母中合成100多种酶组合，并将其新的SL产品鉴定并纯化用于结构研究。最终，将检查这些新SLS的抗菌活动。第三，由于我们不完全了解NR在植物中的生物合成方式，部分原因是我们对NR途径中几种蛋白质成分的了解差异，因此将通过生物化学和遗传学方法来鉴定NR生物合成的其他必要蛋白质。新确定的蛋白质成分可用于设计可在热带para-rubber树之外生产NR的替代生物系统，从而确保聚合物的可持续性。拟议的研究总的来说，将为我们社会中迫切需要的类吸收产品的可持续供应奠定知识的基础。**