SBIR Phase I: Production of high-value chemicals using industrial yeast hosts

这个小型企业创新研究(SBIR)第一阶段项目建议为可扩展的工业酵母宿主开发代谢工程和合成生物学工具包。目前，绝大多数合成生物学工具都是针对工程大肠杆菌或酿酒酵母的，这是一种通常不太适合工业发酵的实验室模型生物。此外，对于那些非常适合工业发酵的宿主来说，还缺乏可用的合成生物学工具。这项研究通过开发一套基本的合成生物学工具来解决这个问题，该合成生物学工具是一种工业上易于处理但研究不足的酵母菌株。研究目标包括构建和鉴定一系列促进遗传物质转移到宿主细胞的表达载体，构建旨在扰乱宿主新陈代谢并将碳流重新导向目标小分子生产的基因文库，以及展示一种减少竞争代谢途径表达的方法。这些工具的原理验证应用将用于展示在工程酵母中丙二酸生物合成的改进。拟议项目的更广泛/商业影响，如果成功，将是能够实现基因改造和设计出强大的工业酵母宿主的技术，消除传统上阻碍商业和学术研究的重大技术障碍。此外，工业酵母宿主基因工具包可能会加快总市值超过300亿美元的一系列生化产品的研发，并提高其商业经济性。目前，这些产品中的绝大多数是通过石化方式生产的，但如果它们可以通过生物方式生产，则存在潜在的成本和环境优势。这项技术将首先应用于丙二酸的商业化，丙二酸是一种高价值的特种化学品，目前从石化中提取。

This Small Business Innovation Research (SBIR) Phase I project proposes to develop a metabolic engineering and synthetic biology toolkit for a scalable, industrial yeast host. Currently, the vast majority of synthetic biology tools are directed toward engineering E. coli or S. cerevisiae, model laboratory organisms that are often poorly suited for industrial fermentations. Furthermore, there is an absence of available synthetic biology tools for those hosts that are well suited for industrial fermentations. This research addresses this problem through the development of a foundational set of synthetic biology tools in an industrially tractable, but under researched yeast strain. The research objectives include construction and characterization of a series of expression vectors that facilitate transfer of genetic material into host cells, construction of a genetic library designed to perturb host metabolism and redirect carbon flux toward production of target small-molecules, and demonstration of an approach to reduce expression of competing metabolic pathways. Proof-of-principle application of the tools will be used to demonstrate improvements in malonic acid biosynthesis in engineered yeast.The broader/commercial impacts of the proposed project, if successful, will be technology that enables genetic modification and engineering of a robust, industrial yeast host, removing significant technical barriers that have traditionally inhibited both commercial and academic research. In addition, the industrial yeast host genetic toolkit may accelerate research and development on, and improve the commercial economics of, a range bio-chemicals with over $30B in aggregate market value. The vast majority of these products are currently produced petrochemically, but there are potential cost and environmental advantages if they can be produced biologically. The technology will first be applied to commercialize malonic acid, a high-value specialty chemical currently derived petrochemically.