SBIR Phase II: Large-scale, high-throughput optimization of gene expression in industrial yeast for improved small molecule production

这个小企业创新研究（SBIR）第二阶段项目的更广泛的影响/商业潜力是开发一种将低价值糖转化为高价值化学品的微生物技术。今天生产的大多数工业化学品都来自石油和其他不可再生的原料。化学工业的长期增长和可持续性受益于开发使用可再生原料的现有化学品的新路线。此外，由于更高的基础设施成本和更严格的环境要求，许多曾经在美国生产的化学品现在都在国外生产。这导致了美国的贸易逆差。第二阶段的提案旨在开发一种发酵技术，将国内种植的农业材料（例如玉米和农业废弃物）转化为高价值的化学品。优化后的发酵工艺在规模上与现有的石化工艺相比具有成本竞争力。如果成功的话，将以300亿美元规模的有机酸市场为目标，促进国内生物化学制造业的发展。该SBIR二期项目旨在开发大规模、高通量的技术来优化工业酵母的基因表达。工业生物技术领域的一个重要问题是设计和优化非学术或模式微生物的发酵性能的能力。大多数分子代谢工程工具都是针对大肠杆菌和酿酒葡萄球菌这两种模型原核和真核微生物开发的，并不适合用于工业相关的微生物。没有这些工具，新的发酵技术的商业化是昂贵和缓慢的。该二期项目的目标是开发和实施一套为耐酸酵母设计的分子生物学工具，并致力于将其应用于改善小分子生产。具体来说，分子生物学工具对于调整（上调或下调）用户定义的基因转录和翻译是有用的。在小规模发酵中，对含有所需基因修饰的工程微生物进行分析，以改进糖的小分子生产。成功的基因改造是那些能够更有效地从糖中生成小分子产品，并理想地减少糖的生物质生成，从而在规模化的商业过程中提供更低的生产成本。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is development of a microbial technology for the conversion of low-value sugars into high-value chemicals. Most industrial chemicals produced today are derived from petroleum and other nonrenewable raw materials. The long-term growth and sustainability of the chemical industry benefits from development of new routes to existing chemicals using renewable raw materials. Furthermore, due to higher infrastructure costs and stricter environmental requirements, many chemicals that were once produced in the United States are now produced abroad. This contributes to the U.S. trade deficit. This Phase II proposal aims to develop a fermentation technology where domestically grown agricultural materials (for example, corn and waste agricultural residues) are converted into high-value chemicals. The optimized fermentation process is estimated to be cost-competitive with the incumbent petrochemical route when scaled. If successful, this proposal will facilitate growth of a domestic bio-chemical manufacturing industry, targeting the $30 billion organic acids market.This SBIR Phase II project proposes to develop large-scale, high-throughput techniques to optimize gene expression in industrial yeast. A significant problem within the field of industrial biotechnology is the ability to engineer and optimize the fermentation performance of non-academic or model microbes. Most molecular metabolic engineering tools are developed for use in two model prokaryotic and eukaryotic microbes, E. coli and S. cerevisiae, and are not suitable for use with industrially relevant microbes. Without these tools it is costly and slow to commercialize new fermentation technologies. The goal of this Phase II project is to develop and implement a set of molecular biology tools designed for acid-tolerant yeast, and working to apply them toward improving small molecule production. Specifically, the molecular biology tools are useful for tuning (up- or down-regulation) user-defined gene transcription and translation. Engineered microbes harboring the desired genetic modification(s) are assayed for improved small molecule production from sugar in small scale fermentations. Successful genetic modifications are those that result in more efficient small molecule product formation from sugar, and ideally decreased biomass formation from sugar, providing a lower production cost in a scaled, commercial process.