Engineering synthetic C1 utilisation in non-conventional organisms for sustainable bioproduction

到目前为止，气候变化可能是对人类持续福祉的最严重威胁。要实现真正可持续的未来，迫切需要实用的技术。一方面，工业排放的碳和农业排放的甲烷是气候变化的两个关键驱动因素。另一方面，石化行业本身固有的不可持续发展是循环经济的一大障碍。已经做出了重大努力来设计微生物系统，以使用生物系统来生产有机化学品，然而这些系统通常依赖于糖原料。通过电还原成甲酸，二氧化碳和甲烷可以简单地转化为微生物原料。这种C1化合物可以被营养格式化的微生物消耗，不幸的是，这些微生物很难通过工程来制造有用的产品。现在，由于合成生物学和新陈代谢工程的发展，尽管具有挑战性，但通过工程改造非格式化营养的工业有机体将甲酸盐用作碳和能源是可能的。在这个项目中，非常规的工业酵母菌将被改造成既能利用甲酸盐作为底物，又能生产高价值产品。这将通过组合合成生物学工具(金门、CRISPR、进化工程、代谢模型)来实现。在这种菌种设计方法中，它有望产生能够1)耐受高浓度甲酸盐的改良菌株2)在其生物量中加入C1底物3)产生细胞能量和4)产生大量工业相关产品。此外，该项目将研究基于甲酸盐的生物过程中细胞间的可变性，这是微生物生物技术目前面临的挑战之一。

Climate change is likely the most serious threat thus far to the continued wellbeing of humanity. Practical, applicable technologies are desperately needed to realise a truly sustainable future.On the one hand, carbon emissions from industry and methane emissions from agriculture are two key drivers of climate change. On the other hand, the inherent unsustainability of the petrochemical industry itself is a major roadblock to a circular economy. Significant efforts have been made to design microbiological systems to manufacture organic chemicals using biological systems, however these often rely on sugar feedstocks. Carbon dioxide and methane can be simply converted into microbial feedstock through electrical reduction into formic acid. This C1 compound can be consumed by formatotrophic microbes, which unfortunately are difficult to engineer to make useful products. Now, thanks to the development of synthetic biology and metabolic engineering, it is possible, though challenging, to engineer non-formatotrophic, industrial organisms to utilise formate as source of carbon and energy. In this project, the non-conventional, industrial yeast Yarrowia lipolytica will be engineered to both utilise formate as substrate and produce high-value products. This will be achieved by a combination of synthetic biology tools (Golden Gate, CRISPR, evolution engineering, metabolic models). In this strain design approach, it is expected to generate improved strains able to 1) tolerate high concentrations of formate 2) incorporate C1 substrates in their biomass 3) generate cellular energy and 4) produce high amount of industrially relevant products. In addition, the project will look into cell-to-cell variability in formate-based bioprocess, which is one of the current challenges of microbial biotechnology.