Using microbial co-cultures involving Yarrowia lipolytica to produce sesquiterpenoids

该项目的中心目标是利用各种生物共培养工程和相关技术在解脂雅罗维菌酵母中实现高产量和工业相关的倍半萜类化合物的生产。倍半萜类化合物是一类高价值的萜类化合物，在医学上具有多种用途，如治疗癌症、神经退行性疾病、炎症、伤口愈合等。解脂亚罗维菌是一种潜在的高产量萜类化合物。不同微生物物种的合作以及单一物种群落内的专业化是提高微生物生物技术生产率的新兴代谢工程战略。我的项目包含几个步骤，以实现基于社区的有效生产倍半萜：1)使用蓝藻或藻类工程分泌蔗糖与解脂亚罗维菌群落喂养它，同时消耗亚罗维菌的废物，工程师解脂亚罗维亚通过基因改造代谢过量生产倍半萜。主要的策略将是过度生产途径酶，导致最终产物，并抑制其他途径分支。3)建立单一物种的Yarrowia群落。Yarrowia生产的柠檬酸是一种废物，大大减少了对所需产品的碳使用量。然而，Yarrowia也能够利用柠檬酸。我的目标是改造这两个菌株，以生产最终的倍半萜产品，但其中一个Yarrowia菌株会消耗柠檬酸废物。4)将第一点和第二点结合到蓝藻-酵母胞内相互作用中。蓝藻会将蔗糖和倍半萜中间体释放到Yarrowia胞浆中，转化为最终产品。5)探索的其他途径：--社区行为的计算模型和现有模型的应用--构建自动定向进化设备，以筛选表现最好的社区，并允许那些社区进一步适应--分析社区成员细胞内发生的事情--将共培养嵌入生物材料中，在伤口中产生与医学相关的化合物

The central aim of this project is utilising various biological co-culture engineering and related techniques to achieve high and industrially relevant production of sesquiterpenoids in the yeast Yarrowia lipolytica. Sesquiterpenoids are types of high-value terpenoids with various uses in medicine such as in the treatment of cancer, neurodegenerative diseases, inflammation, wound healing and others. Yarrowia lipolytica has been identified as a potential high producer of terpenoids. Cooperation of different microbial species as well as specialisation within the single-species community are emerging metabolic engineering strategies to increase the biotechnological productivity of microbes. My project contains several steps towards the efficient community-based production of sesquiterpenoids: 1) Use cyanobacteria or algae engineered to secrete sucrose in a community with Yarrowia lipolytica to feed it while consuming a waste product of Yarrowia, CO22) Engineer Yarrowia lipolytica metabolism by genetic modifications to overproduce sesquiterpenoids. The major strategy would be overproducing pathway enzymes leading to the final product and repressing the other pathway branches. 3) Creating a single-species Yarrowia community. Yarrowia produces citric acid as a waste product which decreases the carbon use towards the desired product significantly. However, Yarrowia is also able to utilise citric acid. I aim to engineer both strains to produce the final sesquiterpenoid product but one of the Yarrowia strains consuming the citric acid waste. 4) I would aim to combine the 1st and 2nd point into cyanobacteria-yeast intracellular interaction. Cyanobacteria would be releasing sucrose and sesquiterpenoid intermediates into Yarrowia cytosol to get converted into the final products. 5) Additional avenues to explore: - Computational modelling of community behaviour and application of existing models- Constructing automated directed evolution devices to screen the best performing communities and allow those to adapt further- Analysis of what happens inside the community member cells- Embedding the co-cultures into biomaterials to produce medically-relevant compounds into wounds