Development of advanced biocatalyst tools and resources to enable biogas-based biomanufacturing.

甲烷营养细菌对富含甲烷的气流进行生物转化，包括厌氧消化产生的沼气，是一条很有希望的途径，可以在评估这些丰富的、浪费的碳源的同时减少温室气体排放。该项目将开发基因工程工具和代谢模型，以推进以甲烷为基础的生物技术，以封存/利用温室气体作为生产可再生燃料和化学品的原料。该项目还将促进北得克萨斯大学对代表不足的学生的培训，这是一所为拉美裔服务的机构，以促进科学劳动力的多样化。此外，该项目将建立一个由不同的高中生、本科生和研究生组成的国际基因工程机器(IGEM)团队，该团队将通过科学推广活动与更广泛的社区接触，促进合成生物学和代谢工程。这项研究的长期目标是开发以甲烷为基础的甲烷和二氧化碳温室气体缓解和转化生物技术。为此，该项目的目标是：1)将胶囊甲基球菌装载到能源部的敏捷生物铸造厂，并开发基因工具，使这种细菌的高通量代谢工程成为可能；2)通过13C通量组学定位和比较胶囊霉菌CH4和二氧化碳在野生型和基因工程菌中的碳通量；以及3)迭代开发预测代谢模型，以指导基于设计-构建-测试-学习的甲烷营养生物催化剂的代谢工程方法。先进的基因工程工具箱的扩展和协调代谢途径的确定调解甲烷/二氧化碳的双重利用和转化，将使这些生物的合理代谢工程能够从单一的碳温室气体中生物制造绿色燃料和化学品。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Biological conversion of methane-rich gas streams, including anaerobic digestion-derived biogas, by methanotrophic bacteria represents a promising route to valorize these abundant, squandered carbon sources while simultaneously mitigating greenhouse gas emissions. This project will develop genetic engineering tools and metabolic models that will advance methanotroph-based biotechnologies aimed at the sequestration/utilization of greenhouse gases as feedstocks for the production of renewable fuels and chemicals. This project will also facilitate the training of underrepresented students at University of North Texas, a Hispanic-serving institution, to promote diversification of the scientific workforce. Further, this project will establish an International Genetically Engineered Machine (iGEM) team consisting of diverse high school, undergraduate, and graduate students that will engage with the broader community to promote synthetic biology and metabolic engineering via scientific outreach activities.The long-term goal of this research is to develop methanotroph-based CH4 and CO2 greenhouse gas mitigation and conversion biotechnologies. To this end, the goals of this project are to 1) onboard Methylococcus capsulatus to the Department of Energy’s Agile Biofoundry and develop genetic tools that enable high-throughput metabolic engineering of this bacterium; 2) map and compare M. capsulatus CH4 and CO2 carbon flux in wild-type and genetically engineered bacteria via 13C fluxomics; and 3) iteratively develop a predictive metabolic model to guide Design-Build-Test-Learn-based metabolic engineering approaches of methanotrophic biocatalysts. Expansion of an advanced genetic engineering toolbox and identification of the coordinated metabolic pathways mediating dual CH4/CO2 utilization and conversion in M. capsulatus will enable the rational metabolic engineering of these organisms for biomanufacturing of green fuels and chemicals from single carbon greenhouse gases.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.