灵芝酸是药用高等真菌灵芝特有的次生代谢产物。其中，灵芝酸单体T和Me具有良好的抗肿瘤及抑制肿瘤转移的活性。然而，缺乏大量、有效的供给是阻碍其广泛应用以及临床前的深入研究的瓶颈。我们的前期工作鉴定获得了数个参与合成灵芝酸单体的关键酶基因。本研究基于以往的工作积累及文献报道的信息，首先构建高产灵芝酸单体T和Me的灵芝基因工程菌株，再就工程菌和野生菌的发酵进行组学（比较转录谱、蛋白质谱和代谢物谱）分析，揭示工程菌株中灵芝酸合成的规律和受调控机制，挖掘菌株代谢改造的新靶点，进而开展工程菌的再改造。最后，进行优化的工程菌（细胞工厂）在反应器培养环境的适配性研究。通过有机结合代谢工程、组学及生物反应器技术，提出灵芝酸单体高效生物合成的新思路新方法，也为其他微生物（特别是高等真菌）次生代谢产物的发酵高产研发提供借鉴和范例。

Ganoderic acids (GAs) are unique secondary metabolites of medicinal mushroom Ganoderma lucidum, of which GA-T and GA-Me were proved to have significant anti-tumor and anti-metastasis activities. However, the lack of substantial and efficient supply of these individual GAs has hindered their widespread application and large-scale pre-clinical studies. A couple of key enzymes, responsible for the biosynthesis of these GAs, were identified in our recent research. Based on the accumulation of our previous long-term related work as well as literature information, in this proposal, an engineered strain of G. lucidum will be constructed as the first step, which could produce more GA-T and GA-Me compared to the wild type. Then, detailed omics analyses (including transcriptomics, proteomics and metabolomics) in fermentation of the engineered strain vs. the wild type will be conducted, so as to clarify the affecting law and acting mechanism of GA biosynthesis in the engineered strain and to find out new potential targets for further metabolic engineering. After the engineered cell factory is re-improved by metabolic engineering, its adaptability to the bioreactor cultivation environment will be investigated. In short, by effectively combining metabolic engineering, omics and bioreactor engineering research, new ideas and new methodologies for reaching hyper-production of these individual GAs will be proposed, and a large-scale highly-efficient production of GA-T and GA-Me is to be achieved. The work will provide a typical example and insights to studies on secondary metabolite production by other microorganisms (especially higher fungi) from strain improvement to fermentation technology innovation.