Methanogenic degradation of branched alkanes in enrichment cultures of production water from a high-temperature petroleum reservoir
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Branched alkanes are important constituents of crude oil and are usually regarded as resistant to microbial degradation, resulting in little knowledge of biochemical processes involved in anaerobic branched alkanes biodegradation. Here, we initiated an incubation study by amendment of iso-C9 (2-methyl, 3-methyl, and 4-methyloctane) as substrates for methanogenic degradation in production water from a high-temperature petroleum reservoir. Over an incubation period of 367 days, significant methanogenesis was observed in samples amended with these branched alkanes. The strong methanogenic activity only observed in iso-C9 amendments suggested the presence of microbial transformation from iso-alkanes into methane. GC-MS-based examination of the original production water identified an intermediate tentatively to be iso-C9-like alkylsuccinate, but was not detected in the enrichment cultures, combined with the successful amplification of assA functional gene in inoculating samples, revealing the ability of anaerobic biodegradation of iso-C9 via fumarate addition pathway. Microorganisms affiliated with members of the Firmicutes, Synergistetes, and methanogens of genus Methanothermobacter spp. were highly enriched in samples amended with iso-C9. The co-occurrence of known syntrophic acetate oxidizers Thermoacetogenium spp. and Methanothermobacter spp. (known hydrogenotrophic methanogens) indicates a potential syntrophic acetate oxidation associated with the methanogenic biodegradation of iso-C9. These results provide some useful information on the potential biodegradation of branched alkanes via methanogenesis and also suggest that branched alkanes are likely activated via fumarate addition in high-temperature petroleum reservoirs.
KeywordsAlkylsuccinate iso-Alkanes or branched alkanes High-temperature petroleum reservoir Methanogenic degradation assA gene
The authors are grateful to the management of Shengli Oilfield for sampling support.
This work was supported by the National Nature Science Foundation of China (Grants No. 41530318, 41373070), NSFC/RGC Joint Research Fund (No. 41161160560), Shanghai Fundamental Research Program (15JC1401400), and the Fundamental Research Funds for the Central Universities (No. 222201817017).
Compliance with ethical standards
This article does not contain any studies with human participants or animals by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
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