Metabolic and Biosynthetic Engineering of Statin-Producing Filamentous Fungus

描述(由申请人提供)： 天然产物是农业和医药用生物活性化合物的极其重要的来源。许多最畅销的药物是天然产品或天然产品先导的半合成衍生品。丝状真菌是人类已知的一些最重要的药物的多产生产者，包括来自诺氏青霉的青霉素和来自曲霉的他汀类药物。因此，从科学和生物技术的角度来看，对具有重要工业意义的丝状真菌进行代谢工程都是一个吸引人的研究目标。在这项工作中，我们将通过代谢和生物合成工程从土曲霉中生产重磅炸弹药物辛伐他汀(Zocor)。辛伐他汀在降低胆固醇水平方面非常有效，是世界上最畅销的药物之一，年销售额超过50亿美元。辛伐他汀是一种源于天然产物洛伐他汀(Mevacor)的半合成药物，目前采用低效且费力的多步骤化学合成方法生产。因此，能够提高辛伐他汀合成效率、降低生产成本的新方法将具有重大意义。在这个项目中，我们将对洛伐他汀生产菌A.terreus进行代谢工程，以直接生物合成辛伐他汀。在广泛的初步成果的基础上，将追求三个具体目标： 目的1：利用土曲霉的代谢工程进行前体定向生物合成。 目的2：前体导向生物合成辛伐他汀的工艺优化。 目的3：土曲霉生物合成途径工程。 这项拟议的研究具有创新性，因为针对辛伐他汀等重磅炸弹药物的生物合成的代谢工程尚未完成。这项工作将是将代谢工程和生物合成工程应用于合成像辛伐他汀一样具有重要商业意义的化合物方面的一个重要里程碑。该项目的成功结果将导致一种全新的辛伐他汀生产方法，该方法比目前的工艺更有效。除了应用驱动的目标，这个项目的结果将有助于我们对Terreus新陈代谢、蛋白质结构和功能以及真菌生物合成途径的基本了解。

DESCRIPTION (provided by applicant): Natural products are extremely important sources of bioactive compounds for agricultural and pharmaceutical applications. Many of the top selling drugs are natural products or semisynthetic derivatives of natural product leads. Filamentous fungi constitutes a prolific producer of some of the most important drugs known to human kind, including penicillin from Penicillium notatum and the statins from Aspergillus species. Metabolic engineering of industrially important filamentous fungi is therefore an attractive research goal from both scientific and biotechnological perspectives. In this work, we will metabolically and biosynthetically engineer the production of the blockbuster drug simvastatin (Zocor) from Aspergillus terreus. Simvastatin is highly effective in lowering cholesterol levels and is one of the best selling drugs in the world with annual sales exceeding $5 billion. Simvastatin is a semisynthetic drug derived from the natural product lovastatin (Mevacor) and is currently produced using an inefficient and laborious multistep chemical synthesis. Therefore, new methods that can increase the efficiency of simvastatin synthesis, and decrease the cost of production will be of significant impact. In this project, we will perform metabolic engineering of the lovastatin producer A. terreus towards direct biosynthesis of simvastatin. Based on extensive preliminary results, three specific aims will be pursued: Aim 1: Metabolic Engineering of A. terreus for Precursor-Directed Biosynthesis. Aim 2: Optimization of the Precursor-Directed Biosynthesis of Simvastatin. Aim 3: Biosynthetic Pathway Engineering of A. terreus. The proposed research is innovative because metabolic engineering towards the biosynthesis of a blockbuster drug such as simvastatin has not been accomplished. This work will represent an important milestone in applying metabolic engineering and biosynthetic engineering towards the synthesis of a compound as commercially important as simvastatin. Successful outcome of this project will lead to a completely new method of producing simvastatin that is more efficient than the current processes. In addition to the application-driven goal, results from this project will contribute to our fundamental understanding of A. terreus metabolism, protein structure and function, as well as fungal biosynthetic pathways.