SBIR Phase I: Broadband focusing for non-invasive cell metabolomics

SBIR 第一阶段：宽带聚焦非侵入性细胞代谢组学

• 批准号: 2221721
• 负责人: Adam Hanninen
• 金额: $ 27.5万
• 依托单位: TRESTLE OPTICS 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: 2023-05-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2221721&HistoricalAwards=false
• 关键词: SBIR; Phase; Broadband; focusing; non

The broader impact of this Small Business Innovation Research (SBIR) Phase I project is aimed toward advancing sustainable production of chemicals using synthetic biology. Here, single-cell microbes are engineered to produce valuable metabolites using enzymes rather than sourcing these chemicals from petroleum. Developing genetically engineered cell strains with high yield remains an ongoing effort due to the complexities in how genetic code leads to phenotype expression. This problem is addressed using a bottom-up approach to screen microbe populations at the single-cell level. The method deployed to identify metabolite content in individual cells is based on infrared (IR)-absorption spectroscopy which is label-free, quantitative, and non-destructive. Synthetic biology is poised to disrupt the chemical value-chain by providing an alternative to petroleum-based chemicals that is sustainable and carbon-neutral. Once a highly productive cell is identified it can be selectively propagated to create enriched cell lines. Innovations in optical microscopy are required to improve the performance of the cell screening instruments, which will allow high-resolution focusing across a broad spectral range. The upgraded platform will optimize yield more quickly, providing value by reducing the upfront cost to develop new industrial cell strains. The proposed project emphasizes optical engineering to develop a microscope designed for high-resolution chemical imaging based on molecular vibrational IR-absorptions. This is achieved by deploying focusing elements that operate over a broad spectral range that extend standard optical microscopes to include mid-infrared light sources. The optical instrument will be used to evaluate chemical content in industrial microbe strains and develop enriched cell lines. These single-cell microbe populations are engineered to produce enzymes used to catalyze the synthesis of valuable metabolites. Yields from individual cells, however, are variable due to genetic mutations in the population. Therefore, a quantitative analytical tool based on IR-spectroscopy that can non-destructively identify highly productive cells for selective propagation is extremely desirable. This bottom-up approach for metabolomic cell screening and directed evolution is an innovation as it is label-free, non-invasive, and has strong chemical specificity. In this project, the team will study an industrial microalgae strain used as a low-cost feedstock supplement and identify cells rich in protein content to enhance the overall protein yield.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.

该小企业创新研究 (SBIR) 第一阶段项目的更广泛影响旨在利用合成生物学推进化学品的可持续生产。在这里，单细胞微生物被设计为利用酶生产有价值的代谢物，而不是从石油中采购这些化学品。由于遗传密码如何导致表型表达的复杂性，开发高产量的基因工程细胞株仍然是一项持续的努力。这个问题可以通过自下而上的方法在单细胞水平上筛选微生物群体来解决。用于识别单个细胞中代谢物含量的方法基于红外 (IR) 吸收光谱，该光谱是无标记、定量和非破坏性的。 合成生物学有望通过提供可持续且碳中性的石油化学品替代品来颠覆化学价值链。 一旦鉴定出高产细胞，就可以选择性地增殖它以创建富集的细胞系。需要光学显微镜的创新来提高细胞筛选仪器的性能，这将允许在广泛的光谱范围内进行高分辨率聚焦。升级后的平台将更快地优化产量，通过降低开发新工业细胞株的前期成本来提供价值。拟议的项目强调光学工程，开发一种基于分子振动红外吸收的高分辨率化学成像显微镜。这是通过部署在宽光谱范围内工作的聚焦元件来实现的，这些元件将标准光学显微镜扩展到包括中红外光源。该光学仪器将用于评估工业微生物菌株中的化学含量并开发富集的细胞系。这些单细胞微生物群体经过改造，可以生产用于催化有价值代谢物合成的酶。然而，由于群体中的基因突变，单个细胞的产量是可变的。因此，非常需要一种基于红外光谱的定量分析工具，能够无损地识别高产细胞以进行选择性繁殖。这种自下而上的代谢组细胞筛选和定向进化方法是一项创新，因为它是无标记、非侵入性的，并且具有很强的化学特异性。在该项目中，该团队将研究一种用作低成本原料补充剂的工业微藻菌株，并鉴定蛋白质含量丰富的细胞，以提高总体蛋白质产量。该奖项反映了 NSF 的法定使命，并通过使用评估结果被认为值得支持。基金会的智力价值和更广泛的影响审查标准。