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

这项小企业创新研究（SBIR）第一阶段项目的更广泛影响旨在利用合成生物学促进化学品的可持续生产。在这里，单细胞微生物被改造成利用酶产生有价值的代谢物，而不是从石油中获取这些化学物质。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个持续的努力。使用自下而上的方法在单细胞水平上筛选微生物种群来解决这个问题。用于鉴定单个细胞中代谢物含量的方法基于红外（IR）吸收光谱，该方法无标记，定量且非破坏性。通过提供一种可持续的、碳中和的石油基化学品替代品，合成生物学有望颠覆化工价值链。一旦一个高产的细胞被鉴定出来，它就可以选择性地繁殖以产生富集的细胞系。光学显微镜的创新需要提高细胞筛选仪器的性能，这将允许在宽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，通过降低开发新的工业细胞株的前期成本来提供价值。本项目以光学工程为重点，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。这是通过部署聚焦元件来实现的，该聚焦元件在宽光谱范围内工作，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价工业微生物菌株的化学成分和培养富集细胞系。这些单细胞微生物种群经过改造，产生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，一种基于红外光谱的定量分析工具，可以非破坏性地识别选择性繁殖的高产细胞是非常需要的。这种自下而上的方法用于代谢组细胞筛选和定向进化是一项创新，因为它无标签，无创，并且具有很强的化学特异性。在这个项目中，该团队将研究一种工业微藻菌株，作为一种低成本的原料补充，并鉴定富含蛋白质含量的细胞，以提高整体蛋白质产量。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.