Methane Oxidation by Salt-Tolerant Zodletone Enrichment

Methane stands out as one of the dominant greenhouse gases in the atmosphere, playing a significant role in global warming. The global warming potency of methane is approximately 28-36 times higher than CO2 and contributes to about 20 percent of the overall warming effect. This study delves into a less-explored methane source the abandoned oil and gas (AOG) wells and produced water storage tanks (PWST). The characteristic elevated salinity of AOGs poses a challenge for microorganisms engaged in methane oxidation. Therefore, this study aims to identify and describe the microbes capable of methane oxidation in saline environments, with potential applications in mitigating methane emissions from AOG wells and PWSTs.We enriched the methane oxidation microbial community by repeatedly feeding methane to Zodletone sediment incubated with Mineral Salts medium. After 6 months of incubation, we extracted total genomic DNA from the enrichment culture using the DNeasy PowerSoil Pro Kit. Shotgun metagenome sequencing was performed (Novogen Corporation) to determine community structure and function of the enrichment. The genomic DNA was sheared into short fragments which were then end-paired, A-tailed and then ligated with Illumina adapter. The fragments with adapters were PCR amplified, size selected, purified and library was prepared. The library was sequenced on Illumina platform and after cleanup, 20,063 reads with an average length of 3,726 bp were obtained