Molecular identification of the transcriptional activator of carotenoid biosynthesis in the fungus Fusarium fujikuroi

真菌藤黑镰刀菌类胡萝卜素生物合成转录激活因子的分子鉴定

• 批准号: 281458954
• 负责人: Dr. Steffen Nordzieke
• 金额: --
• 依托单位: Dep. of Genetics
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/281458954?language=en
• 关键词: Molecular; identification; transcriptional; activator; carotenoid

Carotenoids include over 600 different chemical compounds, bearing a large variety of different characteristics. Many of them are key elements of secondary metabolism of bacteria, plants, and fungi. Due to their individual abilities, they provide a large subset of molecular tasks in protecting individuals against light induced or oxidative damage, which reflects their importance for all classes of eukaryotic life. Animals and humans are unable to produce carotenoids. Nevertheless, they are prerequisites for the formation of Vitamin A, and for the protection of the macula lutea. According to this, humans have to incorporate a sufficient amount of carotenoids through their diets. An undersupply, which is common in developing countries, leads to severe defects in vision, skin composition, and the immune systemFungal systems provide an excellent source for the industrial production of natural carotenoids, thereby avoiding biological inactive isomers which are typical byproducts of chemical synthesis. Additionally, fungi need few requirements, are easy to cultivate and, due to their fermentability, can produce enormous amounts of secondary metabolites. To optimize the industrial production, detailed knowledge about the composition and regulation of fungal carotenoid biosynthesis is important.The plant pathogen Fusarium fujikuroi serves as a fungal model system to study carotenoid biosynthesis since the 1980´s. Due to the already performed research of the host group, the structural genes for all proteins involved in the synthesis of carotenoids are known, whereas the regulation of these genes remains unclear. The aim of this project is to characterize novel regulatory proteins of the carotenoid pathway, such as the predicted CarC, a yet undescribed positive regulator of carotenoid biosynthesis expected to interact with the already described negative regulator CarS. For this, Co Immunoprecipitation experiments, pull-down assays, and a heterologous expression in S. cerevisiae will be utilized, aiming at specific characteristics attributed to CarC. These experiments will lead to a credible list of potential candidates. Genetic deletions of the corresponding genes, followed by in depth analyses of the resulting phenotypes, will serve as control experiments and, simultaneously, provide detailed insights also into potential secondary roles of CarC.Already having a strong background in the field of protein-protein interaction and fungal protein complexes, the applicant is highly qualified to perform the targeted experiments. As the host group is one of the leading facilities in the research on F. fujikuroi and has already characterized the structural genes needed for the carotenoid biosynthesis, the collaboration will close the last scientific gap left in the investigation of the regulation. The obtained results may contribute to optimize the industrial production of carotenoids in fungi, further preventing a Vitamin A undersupply of the population.

类胡萝卜素包括600多种不同的化合物，具有多种不同的特征。它们中的许多都是细菌、植物和真菌次生代谢的关键元素。由于它们的个体能力，它们在保护个体免受光诱导或氧化损伤方面提供了很大的分子任务子集，这反映了它们对所有类别的真核生物的重要性。动物和人类都不能产生类胡萝卜素。然而，它们是形成维生素A和保护黄斑的先决条件。根据这一点，人类必须通过饮食摄入足够量的类胡萝卜素。供应不足在发展中国家很常见，导致视力、皮肤成分和免疫系统严重缺陷。Funga系统为天然类胡萝卜素的工业化生产提供了极好的来源，从而避免了典型的化学合成副产品-生物非活性异构体。此外，真菌需要的条件很少，很容易培养，而且由于它们的发酵性，可以产生大量的次生代谢物。为了优化工业生产，详细了解真菌类胡萝卜素生物合成的组成和调控是很重要的。自20世纪80年代S以来，植物病原菌藤黑镰刀菌一直是研究类胡萝卜素生物合成的真菌模式系统。由于宿主组已经进行的研究，参与类胡萝卜素合成的所有蛋白质的结构基因都是已知的，但这些基因的调控尚不清楚。该项目的目的是表征类胡萝卜素途径的新调节蛋白，例如预测的CarC，这是一种尚未描述的类胡萝卜素生物合成的正调控因子，有望与已描述的负调控因子CARS相互作用。为此，将利用免疫共沉淀实验、下拉试验和在酿酒酵母中的异源表达，针对癌的特定特征。这些实验将产生一份可信的潜在候选人名单。相应基因的遗传删除，随后对产生的表型进行深入分析，将作为对照实验，同时还提供对CarC潜在次要作用的详细见解。申请人已经在蛋白质-蛋白质相互作用和真菌蛋白质复合体领域拥有强大的背景，非常有资格进行有针对性的实验。由于宿主群是Fujikuroi研究的领先机构之一，并且已经确定了类胡萝卜素生物合成所需的结构基因，因此这次合作将弥合在调控研究中留下的最后一个科学空白。所获得的结果可能有助于优化真菌中类胡萝卜素的工业化生产，进一步防止人口维生素A供应不足。