Elucidating and engineering bottromycin biosynthesis

底霉素生物合成的阐明和工程设计

• 批准号: BB/M003140/1
• 负责人: Andrew Truman
• 金额: $ 55.16万
• 依托单位: John Innes Centre
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FM003140%2F1
• 关键词: Elucidating; engineering; bottromycin; biosynthesis

One of the greatest threats to public health in the 21st century is the rise of multi-drug resistant bacterial infections, which has been caused by a shortage in new types of antibiotics, as well as the improper use of antibiotics in medicine and agriculture. This has prompted the World Health Organisation to warn that "the need for action to avert a developing global crisis in health care is increasingly urgent" and the UK's Chief Medical Officer, Prof. Dame Sally Davies, to declare that "we are also not developing new drugs fast enough". One incredibly rich resource for antibiotics are microorganisms that live in soil, and the majority of clinically used antibiotics come from these bacteria. These bacteria have evolved the ability to produce natural products with excellent antibacterial activities, as the ability to kill surrounding bacteria is a big advantage when competing for nutrients.One compound I am currently researching is an antibiotic called bottromycin. The molecule has some highly unusual structural features and is a very active antibiotic towards dangerous infections like the "superbug" MRSA (methicillin-resistant Staphylococcus aureus), which kills tens of thousands of people worldwide each year. Bottromycin has actually been known for many years but problems with its stability and availability have prevented it from being used clinically. However, its entirely novel structure and mode of action make it highly promising antibiotic of the future. An understanding of bottromycin biosynthesis would provide the information necessary for the pathway to be modified to alter the structure of this antibiotic and increase the amount that can be made.Natural products are produced by the action of a series of enzymes (proteins), which are encoded by genes (DNA) in the bacterial genome. I have previously identified the genes required for bottromycin production and will now focus on the details of each biochemical step. I will also develop methods to make new bottromycin-like compounds. From a purely scientific perspective, the biochemical steps in this pathway are highly unusual, and an understanding of the enzyme mechanisms will increase our understanding of enzyme function. The experimental techniques involved in this work include the fermentation of bacterial cultures, the purification of enzymes and the analysis of biochemical reactions using mass spectrometry, which determines the mass of a compound and can provide important structural information.

21世纪对公共卫生的最大威胁之一是耐多药细菌感染的增加，这是由于新型抗生素短缺以及在医学和农业中不当使用抗生素造成的。这促使世界卫生组织发出警告，“采取行动以避免不断发展的全球卫生保健危机的必要性日益迫切”，英国首席医疗官Dame Sally Davies教授宣布“我们开发新药的速度也不够快”。一种非常丰富的抗生素资源是生活在土壤中的微生物，临床上使用的大多数抗生素都来自这些细菌。这些细菌已经进化出了生产具有优异抗菌活性的天然产物的能力，因为在争夺营养物质时，杀死周围细菌的能力是一个很大的优势。我目前正在研究的一种化合物是一种叫做bottromycin的抗生素。这种分子具有一些非常不寻常的结构特征，是一种非常有效的抗生素，可以对抗像“超级细菌”MRSA（耐甲氧西林金黄色葡萄球菌）这样的危险感染，这种细菌每年在全球造成数万人死亡。事实上，人们知道肉霉素已经很多年了，但它的稳定性和可得性问题阻碍了它在临床上的应用。然而，其全新的结构和作用方式使其成为未来非常有前途的抗生素。对botomycin生物合成的理解将提供必要的信息来修改该途径，以改变这种抗生素的结构并增加可以制造的量。天然产物是由细菌基因组中的基因（DNA）编码的一系列酶（蛋白质）的作用产生的。我之前已经确定了肉霉素生产所需的基因，现在将重点放在每个生化步骤的细节上。我还将开发制造新的类似肉霉素的化合物的方法。从纯科学的角度来看，这一途径中的生化步骤是非常不寻常的，对酶机制的理解将增加我们对酶功能的理解。这项工作涉及的实验技术包括细菌培养物的发酵、酶的纯化和使用质谱法分析生化反应，质谱法可以确定化合物的质量并提供重要的结构信息。

项目成果

Dissecting Bottromycin Biosynthesis Using Comparative Untargeted Metabolomics.

• DOI: 10.1002/anie.201604304
• 发表时间: 2016-08-08
• 期刊: ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
• 影响因子: 16.6
• 作者: Crone, William J. K.;Vior, Natalia M.;Santos-Aberturas, Javier;Schmitz, Lukas G.;Leeper, Finian J.;Truman, Andrew W.
• 通讯作者: Truman, Andrew W.

New developments in RiPP discovery, enzymology and engineering.

• DOI: 10.1039/d0np00027b
• 发表时间: 2021-01-01
• 期刊: Natural product reports
• 影响因子: 11.9
• 作者: Montalbán-López M;Scott TA;Ramesh S;Rahman IR;van Heel AJ;Viel JH;Bandarian V;Dittmann E;Genilloud O;Goto Y;Grande Burgos MJ;Hill C;Kim S;Koehnke J;Latham JA;Link AJ;Martínez B;Nair SK;Nicolet Y;Rebuffat S;Sahl HG;Sareen D;Schmidt EW;Schmitt L;Severinov K;Süssmuth RD;Truman AW;Wang H;Weng JK;van Wezel GP;Zhang Q;Zhong J;Piel J;Mitchell DA;Kuipers OP;van der Donk WA
• 通讯作者: van der Donk WA