High methane production by Methanobacterium ferruginis Mic6c05T, a hydrogenotrophic methanogen isolated from a depleted oil and gas field in a neighboring country.

Nusara Yinyom

Department of Microbiology, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand

Kusuma Rintachai

Department of Civil Engineering, Faculty of Engineering, Naresuan University, Phitsanulok, Thailand

Siriwan Wichai

Department of Microbiology, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand

Keywords: methanogenesis, archaea, biogenic gas, methanogenic media, phylogenetic analysis, MPN

Abstract

Some depleted oil and gas fields harbor anaerobic microbes, including methanogens. In this study, methanogens were isolated from a depleted oil and gas field in a neighboring country using enrichment techniques in methanogenic media (ATCC 1340). Five isolates were obtained. Among them, three isolates- ATCC 06.2-1, ATCC 06.2-2, and ATCC reactor 1- demonstrated a strong ability to produce high levels of methane in the ATCC 1340 medium. This culture medium is specifically designed to convert H₂ and CO₂ at a ratio of 4:1, leading to a theoretical total yield of 249.82 µmol of methane. The results indicated that ATCC 06.2-1, ATCC 06.2-2, and ATCC reactor 1 achieved methane production percentages of 94.07%, 75.65%, and 72.05% of the theoretical yield, respectively. Additionally, all three isolates could utilize formate and methanol as substrates for methane production and were classified as hydrogenotrophs. Genetic identification revealed that the three methane-producing isolates- ATCC 06.2-1, ATCC 06.2-2, and ATCC reactor 1- belong to the species Methanobacterium ferruginis Mic6c05^T. The other two isolates, ATCC 03 and ATCC 04.2, were identified as Clostridium sporogenes DSM 795^T. The three methane-producing isolates and the two Clostridium isolates show significant potential for optimizing alternative energy sources.

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