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Demethylated hopanoids in ‘Ca. Methylomirabilis oxyfera’ as biomarkers for environmental nitrite-dependent methane oxidation
Smit, N.T.; Rush, D.; Sahonero Canavesi, D.X.; Verweij, M.; Rasigraf, O.; Guerrero-Cruz, S.; Jetten, M.S.M.; Sinninghe Damsté, J.S; Schouten, S. (2019). Demethylated hopanoids in ‘Ca. Methylomirabilis oxyfera’ as biomarkers for environmental nitrite-dependent methane oxidation. Org. Geochem. 137: 103899. https://dx.doi.org/10.1016/j.orggeochem.2019.07.008
In: Organic Geochemistry. Elsevier: Oxford; New York. ISSN 0146-6380; e-ISSN 1873-5290, meer
Peer reviewed article  

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Author keywords
    Demethylated hopanoids; Candidatus Methylomirabilis oxyfera; Nitrite-dependent methane oxidation; Trisnorhopanoids; 3-Methyl 22,29,30-trisnorhopan-21-one; Methylation

Auteurs  Top 
  • Smit, N.T., meer
  • Rush, D., meer
  • Sahonero Canavesi, D.X.
  • Verweij, M., meer
  • Rasigraf, O.
  • Guerrero-Cruz, S.
  • Jetten, M.S.M.
  • Sinninghe Damsté, J.S, meer
  • Schouten, S., meer

Abstract
    Hopanoids are lipids that are widespread in the bacterial domain and well established molecular biomarkers in modern and paleo environments. In particular, the occurrence of 13C-depleted 3-methylated hopanoids are characteristic of aerobic bacteria involved in methane oxidation. Previously the intra-aerobic methanotroph ‘Candidatus Methylomirabilis oxyfera’ (‘Ca. M. oxyfera’), which performs nitrite-dependent methane oxidation in anoxic environments, has been shown to synthesize bacteriohopanepolyols (BHPs) and their 3-methylated counterparts. However, since ‘Ca. M. oxyfera’ does not utilize methane as a carbon source, its biomass and lipids do not show the characteristic 13C-depletion. Therefore, the detection of ‘Ca. M. oxyfera’ in various environments is challenging, and still underexplored. Here, we re-investigated the hopanoid content of ‘Ca. M. oxyfera’ bacteria using enrichment cultures. We found the GC-amenable hopanoids of ‘Ca. M. oxyfera’ to be dominated by four demethylated hopanoids of which only one, 22,29,30-trisnorhopan-21-one, had been identified previously. The three novel hopanoids were tentatively identified as 22,29,30-trisnorhopan-21-ol, 3-methyl-22,29,30-trisnorhopan-21-one and 3-methyl-22,29,30-trisnorhopan-21-ol. These unique demethylated hopanoids are most likely biosynthesized directly by ‘Ca. M. oxyfera’ bacteria. Bioinformatical analysis of the ‘Ca. M. oxyfera’ genome revealed potential candidate genes responsible for the demethylation of hopanoids. For the sensitive detection of the four trisnorhopanoid biomarkers in environmental samples, a multiple reaction monitoring (MRM) method was developed and used to successfully detect the trisnorhopanoids in a peatland where the presence of ‘Ca. M. oxyfera’ had been confirmed previously by DNA-based analyses. These new biomarkers may be a novel tool to trace nitrite-dependent methane oxidation in various (past) environments.

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