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

这个小企业创新研究（SBIR）第一阶段项目提出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E。coli或S.酿酒酵母，通常不适合工业发酵的模型实验室生物。此外，对于那些非常适合工业发酵的宿主，缺乏可用的合成生物学工具。这项研究通过在工业上易于处理但研究不足的酵母菌株中开发一套基础合成生物学工具来解决这个问题。研究目标包括一系列表达载体的构建和表征，这些载体有助于将遗传物质转移到宿主细胞中，构建一个基因库，该基因库旨在干扰宿主代谢并将碳通量重新定向到目标小分子的生产，并展示一种减少竞争代谢途径表达的方法。该工具的原理验证应用将用于证明工程酵母中丙二酸生物合成的改进。如果成功，拟议项目的更广泛/商业影响将是能够进行基因修饰和工程改造的技术，强大的工业酵母宿主，消除传统上抑制商业和学术研究的重大技术障碍。此外，工业酵母宿主遗传工具包可以加速研究和开发，并提高一系列生物化学品的商业经济性，总市场价值超过300亿美元。目前绝大多数这些产品都是通过石油化学方法生产的，但如果可以通过生物方法生产，则具有潜在的成本和环境优势。该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，目前从石油化工中提取。

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.