SBIR Phase I: Boosting Industrial Bio-Fermentation with Microbial Stem Cells

SBIR 第一阶段：利用微生物干细胞促进工业生物发酵

• 批准号: 2222602
• 负责人: Nikolai Mushnikov
• 金额: $ 27.41万
• 依托单位: ASIMICA LLC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2222602&HistoricalAwards=false
• 关键词: SBIR; Phase; Boosting; Industrial; Bio

The broader impact of this Small Business Innovation Research (SBIR) Phase I project is to reimagine bio-manufacturing with a novel platform technology that could boost the yields of many products, including food additives, biomaterials precursors, biofuels, and pharmaceuticals. The technological advancement addresses a fundamental issue that limits conventional bio-fermentation, which is that producing cells suffer limited health and viability in exchange for higher yields. In this proposal, genetic tools will be used to divide the labor of cell reproduction and product synthesis into two different cell types, called stem cells and factory cells. As older factory cells become exhausted, productivity is maintained by new factory cells, which are born from the stem cell population. The approach may be particularly well suited to biofuels and other molecules that are difficult to produce in large quantities by conventional bio-fermentation because the product is toxic to the cells that make it. It could be applied toward increasing the profitability of existing bio-processes and also for bringing new products to market, which are currently too difficult to produce. In this project, the team seeks to demonstrate the benefits of producing a fuel (limonene) and a dairy enzyme (chymosin), as proof of its application in biofuel and agricultural sectors. Broad industrial implementation will advance bio-manufacturing toward the ‘green’ revolution, contributing to the development of cleaner industries and decreasing US and global reliance on fossil fuels.This project aims to solve two major limitations of microbial fermentation processes: metabolic exhaustion and genetic drift. These are nearly universal problems in the industry. Highly producing cells can become inactive due to the lack of metabolic resources, cytotoxic effects of products, and mutations that break the biosynthetic pathway. In this project, Microbial Stem Cell Technology (MiST) uncouples growth and production by establishing a multicellular system. One cell type is dedicated to product synthesis (factory cells), while another (stem cells) is responsible for cell division and the generation of new factory cells. As older factory cells lose productivity, the bioreactor is continuously replenished with new factory cells, derived from the stem cell population. By maintaining an active factory cell population, MiST-supported cultures are expected to exhibit increased production longevity and higher overall yield than conventional bio-fermentations. This project aims to validate the technology in E. coli engineered to produce limonene, a precursor for biodiesel and other useful chemicals. In the factory cells, T7RNAP will drive high-level expression of a suite of biosynthetic enzymes. Since limonene has a cytotoxic effect on producing cells, MiST-supported factory cell replenishment is expected to increase productivity by more than 2-fold compared to the conventional limonene-producing strains.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这项小型企业创新研究（SBIR）I阶段项目的更广泛影响是通过一种新型的平台技术重新想象生物制造，该技术可以提高许多产品的产量，包括食品添加剂，生物材料前体，生物织物和药品。技术进步解决了一个基本问题，该问题限制了常规的生物发酵，即产生细胞的健康和生存能力有限，以换取更高的收益率。在此提案中，遗传工具将用于将细胞繁殖和产品合成的分为分为两种不同的细胞类型，称为干细胞和工厂细胞。随着较旧的工厂细胞的疲惫，生产力由新的工厂细胞维持，这些新工厂细胞由干细胞种群诞生。该方法可能特别适合生物燃料和其他通过常规生物发酵大量生产的生物燃料和其他分子，因为该产物对生产物的细胞有毒。它可以应用于提高现有生物过程的盈利能力，还可以将新产品推向市场，这些产品目前难以生产。在这个项目中，该团队试图证明生产燃料（柠檬烯）和乳制品（芝麻素）的好处，以证明其在生物燃料和农业领域中的应用。广泛的工业实施将推进生物制造，以实现“绿色”革命，从而促进了清洁行业的发展，并减少了美国和全球对化石燃料的依赖。该项目旨在解决微生物发酵过程的两个主要局限性：代谢精疲力尽和遗传漂移。这些几乎是行业中的普遍问题。由于缺乏代谢资源，产物的细胞毒性作用以及破坏生物合成途径的突变，高度产生的细胞可能变得无活跃。在这个项目中，微生物干细胞技术（MIST）通过建立多细胞系统来取消生长和生产。一种细胞类型专用于产品合成（工厂细胞），而另一种细胞类型则负责细胞分裂和新的工厂细胞的产生。随着较老的工厂细胞失去生产力，生物反应器被从干细胞种群中得出的新工厂细胞不断复制。通过维持活跃的工厂细胞人群，预计被雾化的培养物的生产寿命增加，总产量会比传统的生物发酵量更高。该项目旨在验证旨在生产柠檬烯的大肠杆菌中的技术，这是生物柴油和其他有用化学物质的前体。在工厂细胞中，T7RNAP将驱动一组生物合成酶的高级表达。由于柠檬烯对生产细胞具有细胞毒性作用，因此与传统的产生limononene的菌株相比，预计雾化的工厂细胞补充有望提高生产率超过2倍。这一奖项反映了NSF的法定任务，并被认为是通过基金会的知识分子优点和广泛的影响来评估的法定任务。