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，在酵母中实现高产量和工业相关的倍半萜类产品。倍半萜类化合物是一种高价值萜类化合物，在医学上有多种用途，如治疗癌症、神经退行性疾病、炎症、伤口愈合等。多脂耶氏菌已被确定为一种潜在的萜类化合物高产菌。不同微生物物种的合作以及单物种群落内的专业化是新兴的代谢工程策略，以提高微生物的生物技术生产力。我的项目包含了几个步骤，以有效地以社区为基础生产倍半萜类化合物：1)在一个有多脂耶氏菌的社区中，使用经过改造的蓝藻或藻类分泌蔗糖来喂养它，同时消耗耶氏菌的废物，CO22)通过基因改造来改造多脂耶氏菌的代谢，以过量生产倍半萜类化合物。主要的策略是过量生产导致最终产物的途径酶，并抑制其他途径分支。3)创建单一物种耶氏菌群落。耶氏菌产生柠檬酸作为废物，这大大减少了对所需产品的碳使用。然而，耶氏菌也能够利用柠檬酸。我的目标是设计这两种菌株来生产最终的倍半萜类产物但其中一种耶氏菌消耗柠檬酸废物。4)我的目标是将第一点和第二点结合到蓝藻-酵母细胞内相互作用中。蓝藻会将蔗糖和倍半萜类中间产物释放到耶氏菌细胞质中，转化为最终产物。5)其他探索途径：-社区行为的计算建模和现有模型的应用-构建自动化定向进化设备以筛选表现最佳的社区并允许这些社区进一步适应-分析社区成员细胞内部发生的情况-将共培养物嵌入生物材料中以产生与医学相关的化合物到伤口中

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