SBIR Phase II: Novel Proteolysis-based Tools for Metabolic Engineering

SBIR 第二阶段：基于蛋白水解的新型代谢工程工具

• 批准号: 1256446
• 负责人: Jason Kelly
• 金额: $ 50万
• 依托单位: Ginkgo BioWorks
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-15 至 2019-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1256446&HistoricalAwards=false
• 关键词: SBIR; Phase; II; Novel; Proteolysis

This Small Business Innovation Research (SBIR) project aims to engineer microbes for the cost-effective production of specialty chemicals. Currently, engineered microbial strains bear mutations that increase the production of chemicals of interest by inhibiting the cell's ability to produce off pathway chemicals. These "loss-of-function" mutations are critical as they effectively channel the cell's metabolic flux toward the product of interest. This both boosts the production efficiency and eases downstream purification by eliminating the accumulation of undesirable but chemically-similar contaminants. Unfortunately, these mutations may also decrease the fitness of the cells and, as a result, the growth media must be supplemented with costly nutrients. Technical research herein will assess the feasibility of applying novel regulated proteolysis technology to simultaneously direct maximal metabolic flux toward the target chemical of interest while avoiding the need to supplement the growth media. If successful, this technology would provide a great cost savings and enable fermentative production to be applied more broadly in the production of specialty chemicals.The broader impact/commercial potential of this project is to provide a stable and cost-effective fermentative production route to a specialty chemical. Fermentative production of chemicals offers many advantages over traditional petrochemical or extraction-based production processes. Petrochemical production maintains the nation?s reliance on an unsustainable feedstock (oil) and also leads to national security issues as the US is largely dependent on foreign oil sources. Chemical production via extraction from plant materials also has ecological challenges. The process often uses toxic solvents, and may rely on unsustainable farming practices for many plants that are not traditional food crops. Engineered microbes fermented on sugar feedstock produced using high-efficiency agricultural practices offer a stable alternative for producing specialty chemicals, both in terms of supply and price.

这个小型企业创新研究（SBIR）项目旨在为具有成本效益的特种化学品生产设计微生物。目前，工程微生物菌株承受突变，通过抑制细胞产生通路外化学物质的能力来增加感兴趣的化学物质的产生。这些“功能丧失”突变是至关重要的，因为它们有效地将细胞的代谢通量引向感兴趣的产物。这既提高了生产效率，又通过消除不需要但化学相似的污染物的积累而简化了下游的净化。不幸的是，这些突变也可能降低细胞的适应性，因此，生长培养基必须补充昂贵的营养物质。本文的技术研究将评估应用新型调节蛋白水解技术的可行性，同时将最大代谢通量导向感兴趣的目标化学物质，同时避免需要补充生长介质。如果成功，这项技术将大大节省成本，并使发酵生产更广泛地应用于特种化学品的生产。该项目的更广泛的影响/商业潜力是为特种化学品提供稳定和具有成本效益的发酵生产路线。与传统的石化或萃取生产工艺相比，化学制品的发酵生产具有许多优点。石油化工生产维持国家？美国对一种不可持续的原料（石油）的依赖，也会导致国家安全问题，因为美国在很大程度上依赖外国石油来源。从植物原料中提取的化学品生产也面临生态挑战。该过程通常使用有毒溶剂，并且可能依赖于非传统粮食作物的许多植物的不可持续的耕作方式。在高效农业生产的糖原料上发酵的工程微生物，在供应和价格方面都为生产特种化学品提供了稳定的选择。