Ectoine production by newly isolated halophilic methanotroph Methylotuvimicrobium percolatum using surplus methane from anaerobic digestion

Anaerobic digestion (AD) processes that convert waste such as livestock wastewater into biomethane for energy recovery are expanding globally. However, biogas exceeding storage capacity is typically flared, resulting in the release of carbon dioxide, a greenhouse gas. To address this issue, we focused on halophilic methane-oxidizing bacteria (methanotrophs) capable of converting methane into ectoine, a high-value compatible solute widely used in more than 400 cosmetic active ingredients and valued at approximately US$1,000/kg. Nevertheless, research on halophilic methanotrophs and ectoine biosynthesis remains limited. Here, we report the successful isolation of the Type I methanotroph Methylotuvimicrobium percolatum from activated sludge in a brackish water environment. Complete genome sequencing revealed genes for methane oxidation, osmoadaptation, and notably the ectABCask cluster responsible for ectoine biosynthesis. Activity tests under varying NaCl concentrations confirmed ectoine production, with a maximum yield of 20.1 mg ectoine/g-DCW achieved at 25% methane, 30 {degrees}C, and 3 wt% salinity. Biomass growth remained robust up to 6% salinity but declined sharply at higher NaCl concentrations, indicating that optimal conditions for growth and ectoine synthesis do not entirely coincide. These findings advance the fundamental understanding of halophilic methanotroph physiology and highlight their potential as sustainable platforms for converting surplus methane from AD processes into commercially valuable ectoine.