BioMADE: Driving Cost Reduction in Biomanufacturing Biomaterials from Methane: Engineering Novel Strains to Increase Downstream Processing Efficiency

Methane is a greenhouse gas. It is made in landfills and wastewater treatment facilities. Certain microbes can grow on methane. These microbes can also produce biodegradable polymers. However, separating these polymers from the cells that produce them and then purifying them is difficult and expensive. One promising biodegradable polymer, PHA, is the focus of this project. To reduce the cost of purification, methane-consuming bacteria that produce PHA will be genetically engineered to accomplish two objectives. One is to cause the cells to automatically break open after producing PHA. The second is to reduce the amount of protein produced to simplify purification. This could significantly improve the affordability of PHA. Educational and workforce development programs will prepare students to join the biomanufacturing industry. The overarching goal is to significantly reduce the downstream processing (DSP) costs associated with methane-derived polyhydroxyalkanoate (PHA) production through advanced genetic engineering of Type II methanotrophic bacteria. Two specific bacterial strains will be targeted for these genetic modifications: the well-established model strain Methylosinus trichosporium and the Mango Materials’ proprietary strain (str. mngo). Genetic modifications will target pathways to facilitate easier recovery of PHAs from the fermentation process. The project employs iterative Design-Build-Test-Learn (DBTL) cycles supported by proteomic analyses, techno- economic analysis (TEA), and life cycle assessment (LCA). The goal is a reduction of 5–10% in capital expenditures and up to 15% in operational expenditures. This research will yield robust genetic engineering tools and engineered strains, while also significantly advancing fundamental scientific understanding in synthetic biology, metabolic engineering, and microbial physiology, particularly in non-model organisms that have traditionally been challenging to engineer. A detailed case study and a specialized industry workshop will facilitate the dissemination of a generalizable framework for DSP-focused genetic engineering, directly benefiting the broader biomanufacturing community. This project is being jointly supported by ENG/CBET/CBE and the BioMADE Manufacturing Innovation Institute. 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.