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体流（包括厌氧消化产生的沼气）进行生物转化，是一种有希望的途径，可以在减少温室气体排放的同时，对这些丰富的、被浪费的碳源进行价值转化。该项目将开发基因工程工具和代谢模型，以推进以甲烷氧化菌为基础的生物技术，目的是将温室气体作为生产可再生燃料和化学品的原料进行封存/利用。该项目还将促进北德克萨斯大学（一所服务于西班牙裔的机构）代表性不足的学生的培训，以促进科学工作人员的多样化。此外，该项目将建立一个由不同高中、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合作，促进合成生物学和代谢工程。本研究的长期目标是开发基于甲烷氧化菌的CH4和CO2温室气体减缓和转化生物技术。为此，该项目的目标是：1)将荚膜甲基球菌（Methylococcus capsulatus）运送到能源部的敏捷生物工厂（Agile bifoundry），并开发能够实现这种细菌高通量代谢工程的遗传工具；2)利用13C通量组学技术绘制并比较野生型和基因工程菌中荚膜分枝杆菌CH4和CO2的碳通量；3)迭代开发预测代谢模型，指导基于设计-构建-测试-学习的甲烷化生物催化剂代谢工程方法。扩展先进的基因工程工具箱和确定协同代谢途径介导的双重CH4/CO2的利用和转化，将使这些生物的合理代谢工程从单一碳温室气体的生物制造绿色燃料和化学品。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。

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.