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

卡特莱链霉菌是一种具有产生氟代谢产物的不同寻常的能力的细菌。它们是有毒的化合物氟乙酸酯和氟化氨基酸4-氟苏氨酸(一种抗生素)。近年来，我们已经能够确定导致卡特利氏链霉菌最初氟化事件的酶。该酶能将S-腺苷-L-蛋氨酸和氟离子转化为5‘-氟-5’-脱氧腺苷和L-蛋氨酸。这是一项重要的发现，因为有机氟化合物作为一类化合物，在精细化学品、制药和农用化学品行业中产生了巨大的影响，并且在新医药产品的发现和开发中继续产生越来越大的影响。到目前为止，所有的含氟化合物都是通过化学方法合成的，因此这种可以吸收无机氟并将其转化为有机氟的酶及其相关基因的鉴定，为利用生物技术而不是化学方法生产有机氟化合物提供了新的前景。我们预计，在不久的将来，我们将能够转移氟化酶基因和相关的生物合成和管理基因(例如。氟乙酸酯抗性基因、氟离子吸收基因)进入宿主生物，并在宿主体内“启动”氟代谢产物的产生。在有机氟化学的背景下，这是新的和令人兴奋的。要做到这一点，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后果来完成。最近的进展已经确定了氟化酶基因以及该基因两侧约10个基因的簇。在这个簇中，存在一些但不是全部的氟乙酸酯和4-氟苏氨酸的生物合成途径基因。这项提议的一个主要焦点是识别剩余的基因，探索基因产物(酶/蛋白质)并对其进行表征。我们还将利用这些基因作为进一步基因行走的焦点，探索侧翼基因。这项研究的总体重点是在遗传水平上表征氟代谢产物的生物合成，并评估相关基因的蛋白质产物以及相关酶的功能和机制。这些信息将被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。

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.