Enzymatic fluorination in Streptomyces cattleya. Setting a framework for biotechnological development.

卡特兰链霉菌中的酶促氟化。

• 批准号: BB/F007426/1
• 负责人: David O'Hagan
• 金额: $ 58.37万
• 依托单位: University of St Andrews
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FF007426%2F1
• 关键词: Enzymatic; fluorination; Streptomyces; cattleya.; Setting

Streptomyces cattleya is a bacterium which has the unusual ability to generate fluorometabolites. These are the toxic compound fluoroacetate and the fluorinated amino acid 4-fluorothreonine (an antibiotic). In recent years we have been able to identify the enzyme responsible for the initial fluorination event in S. cattleya. This enzyme converts S-adenosyl-L-methionine (SAM) and fluoride ion to 5'-fluoro-5'-deoxy-adenosine (5'-FDA) and L-methionine. This is an important discovery because organo-fluorine compounds as a class, have made a huge impact in the fine chemicals, pharmaceuticals and agrochemicals industries and they continue to have a growing impact in the discovery and development of new pharmaceutical products. All fluorinated compounds to date are made by chemical methods thus the identification of this enzyme, which can take inorganic fluoride and convert it to organic fluorine, and the genes associated with this enzyme, offer new prospects for the biotechnological, rather than the chemical, production of organo-fluorine compounds. We envisage that in the near future we will be able to move the fluorinase gene and the related biosynthetic and management genes (eg. fluoroacetate resistance gene, fluoride ion up take genes) into host organisms and 'kick start' fluorometabolite production in the hosts. This is novel and exciting in the context of organo-fluorine chemistry. To do this we need to identify as many of the relevant genes as possible and establish their roles. This will be done by gene knockout experiments and exploring the consequences. Recent progress has identified the fluorinase gene as well as a cluster of about 10 genes flanking this gene. In this cluster some, but not all, of the biosynthetic pathway genes to fluoroacetate and 4-fluorothreonine are present. A major focus of this proposal is to identify the remaining genes, explore the gene products (enzymes/proteins) and characterise these. We will also use these genes as the focal point for further gene walking to explore flanking genes. The overall focus of this research is to characterise fluorometabolite biosynthesis at the genetic level and assess the protein products of relevant genes and the function and mechanisms of relevant enzymes. This information will be used to develop biotechnological tools for the production of novel organofluorine compounds by fermentation methods.

卡特兰链霉菌是一种能产生荧光代谢产物的细菌。它们是有毒的化合物氟乙酸盐和氟化氨基酸4-氟代苏氨酸（一种抗生素）。近年来，我们已经能够鉴定出在S.卡特兰该酶将S-腺苷-L-甲硫氨酸（SAM）和氟离子转化为5 '-氟-5'-脱氧腺苷（5 '-FDA）和L-甲硫氨酸。这是一个重要的发现，因为有机氟化合物作为一类化合物，在精细化学品、制药和农用化学品工业中产生了巨大的影响，并且它们在新医药产品的发现和开发中继续产生越来越大的影响。迄今为止，所有氟化化合物都是通过化学方法制备的，因此这种酶的鉴定，可以将无机氟化物转化为有机氟，以及与这种酶相关的基因，为生物技术而不是化学生产有机氟化合物提供了新的前景。我们设想，在不久的将来，我们将能够移动氟化酶基因和相关的生物合成和管理基因（如基因）。氟乙酸抗性基因、氟离子摄取基因）进入宿主生物体，并在宿主中“启动”氟代代谢物的产生。这在有机氟化学的背景下是新颖的和令人兴奋的。为此，我们需要识别尽可能多的相关基因并确定它们的作用。这将通过基因敲除实验和探索结果来完成。最近的进展已经确定了氟酶基因以及该基因侧翼的约10个基因的簇。在这个簇中，存在一些但不是全部的生物合成途径基因，以氟乙酸和4-氟代苏氨酸。该提案的一个主要重点是识别剩余的基因，探索基因产物（酶/蛋白质）并对其进行测序。我们还将使用这些基因作为进一步基因步行的焦点，以探索侧翼基因。本研究的总体重点是在基因水平上研究氟代谢物的生物合成，并评估相关基因的蛋白质产物以及相关酶的功能和机制。这些信息将用于开发生物技术工具，通过发酵方法生产新型有机氟化合物。

项目成果

Fluorinated 5- and 7-membered carbacycle motifs by reaction of difluorocarbene with acetylene ethers.

• DOI: 10.1039/c3cc39066g
• 发表时间: 2013-02
• 期刊: Chemical communications
• 影响因子: 4.9
• 作者: P. W. Chia;D. Bello;A. Slawin;D. O'Hagan

Mechanistic insights into water activation in SAM hydroxide adenosyltransferase (duf-62).

SAM氢氧化物腺基转移酶（DUF-62）中水活化的机械洞察力。

• DOI: 10.1002/cbic.200900369
• 发表时间: 2009-10-12
• 期刊: CHEMBIOCHEM
• 影响因子: 3.2
• 作者: Deng, Hai;McMahon, Stephen A.;Eustaquio, Alessandra S.;Moore, Bradley S.;Naismith, James H.;O'Hagan, David
• 通讯作者: O'Hagan, David

Engineering fluorometabolite production: fluorinase expression in Salinispora tropica Yields Fluorosalinosporamide.

• DOI: 10.1021/np900719u
• 发表时间: 2010-03-26
• 期刊: JOURNAL OF NATURAL PRODUCTS
• 影响因子: 5.1
• 作者: Eustaquio, Alessandra S.;O'Hagan, David;Moore, Bradley S.
• 通讯作者: Moore, Bradley S.