Illuminating and exploiting programmed O-methylation in trans-AT polyketide synthases

阐明和利用反式 AT 聚酮合酶中的程序化 O-甲基化

• 批准号: BB/W003171/1
• 负责人: Józef Lewandowski
• 金额: $ 101.3万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW003171%2F1
• 关键词: Illuminating; exploiting; programmed; methylation; trans

There is an urgent need to produce novel derivatives of natural product-based medicines, such as antibiotics and anticancer agents, to overcome drug resistance. Moreover, many herbicides, insecticides, and fungicides, which play an essential role in the protection of food crops, are natural products and new derivatives with lower toxicity and greater efficacy are needed to feed the burgeoning global population. Development of novel methodologies for structural modification of natural products and enhancing their production levels are important applications of synthetic biology. Many important natural products are built by modular multienzyme assembly lines. These offer strong potential for rational bioengineering to create novel natural product derivatives. However, a lack of detailed molecular insight into the structure, mechanism, and substrate tolerance of these inherently mobile protein machines has limited progress in this endeavour. Thus, there is a need to develop novel structure-based approaches to elucidating the mechanism and substrate specificity determinants of specific components of such machinery. This project aims to develop an atomic-level understanding of how assembly line machinery appends methyl groups to specific oxygen atoms during assembly of an important class of natural products known as polyketides. To do this, we will develop and apply strategies for capturing intrinsically dynamic components of these machines in specific functional states, which will facilitate the determination of their molecular structures using a variety of methods, including nuclear magnetic resonance spectroscopy, carbene-footprinting mass spectrometry and cryo-electron microscopy. We will also investigate the substrate tolerance of key parts of the machinery using chemically synthesised probe molecules. The new knowledge gained will inform future efforts to bioengineer these assembly lines to produce novel natural product derivatives with altered pharmacokinetic and pharmacodynamic profiles. This has the potential to exploited by UK-based pharmaceutical, agrochemical and biotechnology companies actively engaged in the development of natural product-based consumer products, which will ultimately benefit to wider society.

迫切需要生产以天然产物为基础的药物的新型衍生物，如抗生素和抗癌剂，以克服耐药性。此外，许多在保护粮食作物方面发挥重要作用的除草剂、杀虫剂和杀菌剂都是天然产品，需要毒性更低、药效更高的新衍生品来养活迅速增长的全球人口。开发天然产物结构修饰的新方法并提高其生产水平是合成生物学的重要应用。许多重要的天然产物都是由模块化的多酶流水线生产的。这些为合理的生物工程创造新的天然产品衍生品提供了强大的潜力。然而，对这些固有的移动蛋白质机器的结构、机制和底物耐受性缺乏详细的分子洞察力，限制了这一努力的进展。因此，有必要开发新的基于结构的方法来阐明这种机械的特定组件的机制和底物特异性决定因素。该项目旨在对装配线机械如何在组装一类重要的天然产品聚酮的过程中将甲基添加到特定的氧原子进行原子水平的理解。为此，我们将开发和应用策略来捕获这些机器在特定功能状态下的内在动态组件，这将有助于使用各种方法确定其分子结构，包括核磁共振光谱、卡宾足迹质谱仪和冷冻电子显微镜。我们还将使用化学合成的探针分子来研究机械关键部件的衬底耐受性。所获得的新知识将为未来对这些装配线进行生物工程以生产具有改变的药代动力学和药效学特征的新型天然产品衍生物的努力提供信息。总部设在英国的制药、农用化学品和生物技术公司有可能利用这一潜力，积极从事以天然产品为基础的消费产品的开发，最终将使更广泛的社会受益。