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.

这个小企业创新研究（SBIR）第一阶段项目的更广泛影响是利用新颖的平台技术重新构想生物制造，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、生物燃料和药品。解决了限制传统生物发酵的一个基本问题，即生产细胞的健康和活力受到限制，以换取更高的产量。在该提案中，遗传工具将用于将细胞繁殖和产品合成的劳动分为两个不同的部分。当旧的工厂细胞耗尽时，由干细胞群产生的新工厂细胞可以维持生产力。这种方法可能特别适合生物燃料和其他难以生产的分子。通过传统生物发酵大量生产，因为该产品对制造它的细胞有毒，它可以用于提高现有生物工艺的盈利能力，也可以用于将目前难以生产的新产品推向市场。在这个项目中，团队力求展示的好处。生产燃料（柠檬烯）和乳制品酶（凝乳酶），作为其在生物燃料和农业领域应用的证明，将推动生物制造走向“绿色”革命，促进清洁工业的发展并减少美国的能源消耗。该项目旨在解决微生物发酵过程的两个主要限制：代谢衰竭和遗传漂变，这些是该行业几乎普遍存在的问题。的影响在这个项目中，微生物干细胞技术（MiST）通过建立一个多细胞系统来分离生长和生产。一种细胞类型专门用于产品合成（工厂细胞），而另一种细胞类型（干细胞）。负责细胞分裂和新工厂细胞的生成，当旧工厂细胞丧失生产力时，生物反应器会不断补充来自干细胞群的新工厂细胞，MiST 支持的。与传统的生物发酵相比，该培养物预计会表现出更长的生产寿命和更高的总体产量。该项目旨在验证柠檬烯（生物柴油和其他有用化学品的前体）的大肠杆菌技术。T7RNAP 将在工厂细胞中发挥作用。由于柠檬烯对生产细胞具有细胞毒性作用，因此 MiST 支持的工厂细胞补充预计可将生产率提高 2 倍以上。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。