Abstract

This study presents a novel approach to mitigate methane emissions while simultaneously addressing the growing demand for sustainable animal feed through the development of an engineered methanotrophic strain, Methylomicrobium buryatense 5GB1C-RO1. Utilizing advanced genetic engineering techniques, including CRISPR/Cas9 and horizontal gene transfer, we have optimized the ribulose monophosphate (RuMP) cycle and enhanced oxidase activity in this strain. The bioconversion process is facilitated by innovative bioreactor designs, including Two-Phase Partitioning Bioreactors (TPPBs) and Inverse Membrane Bioreactors (IMBRs), which significantly improve methane solubility and mass transfer. Through metabolic flux analysis and computational modeling, we have achieved high biomass yields and efficient methane utilization. The resulting biofeed demonstrates a superior nutritional profile, with optimized macronutrient content and essential components. This integrated approach not only contributes to greenhouse gas mitigation but also offers a promising solution for sustainable animal nutrition. Our findings suggest that the 5GB1C-RO1 strain and associated bioprocesses have the potential to revolutionize both environmental protection and agricultural sustainability