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Permanent link (DOI): https://doi.org/10.7939/R31X0G

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Using "omics" approaches to study anaerobic hydrocarbon biodegradation by microbes indigenous to oil sands tailings ponds Open Access

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Other title
Subject/Keyword
methanogenic hydrocarbon degradation
metatranscriptomics
oil sands tailings ponds
metagenomics
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Tan, Boon-Fei
Supervisor and department
Dr. Julia Foght
Examining committee member and department
Elizabeth Edwards (Engineering and Applied Chemistry, University of Toronto)
Ania Ulrich (Civil and Environmental Engineering)
Rebecca Case (Biological Sciences)
Lisa Stein (Biological Sciences)
Department
Department of Biological Sciences
Specialization
Microbiology and Biotechnology
Date accepted
2014-01-29T16:05:47Z
Graduation date
2014-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
In oil sands tailings ponds, methanogenesis is driven in part by the degradation of hydrocarbons in residual solvents used as a diluent during bitumen extraction, such as naphtha. Alkanes constitute a large proportion of these unrecovered hydrocarbons in mature fine tailings (MFT). Methanogenic degradation of alkanes has been poorly described in the literature. Fumarate addition, widely reported for activation of alkanes and monoaromatic compounds under sulfate- and nitrate-reducing conditions, has not been demonstrated conclusively for alkane degradation under methanogenic conditions because signature metabolites and key organisms have not been detected and/or isolated. In order to understand methanogenic alkane degradation by microorganisms indigenous to oil sands tailings ponds, a model alkane-degrading culture (SCADC) was established using MFT obtained from Mildred Lake Settling Basin (MLSB) tailings pond. SCADC degraded many lower molecular weight alkanes, represented by n-C 6 -C 10, 2-methylpentane and methylcyclopentane during year-long methanogenic incubation, but expected fumarate addition products were only detected for 2-methylpentane and methylcyclopentane. Nucleic acids isolated from SCADC were subjected to metagenomic and metatranscriptomic analysis using Illumina Hi-Seq. Metagenomic binning using multiple approaches recovered several partial genomes, including novel syntrophic Desulfotomaculum and Smithella spp. that are genetically capable of fumarate addition, which was previously unknown. Metatranscriptomic analysis further confirmed the high expression of genes encoding enzymes for alkane addition to fumarate byDesulfotomaculum but not Smithella during active methanogenesis, indicating the importance of Firmicutes in fumarate activation of low molecular weight alkanes. Data mining of metagenomes of MLSB and hydrocarbon-impacted environments recovered novel fumarate addition genes undescribed previously, indicating the overall ubiquitous nature of these genes in anoxic environments. Comparative metagenomic analysis of SCADC to two other metagenomes of methanogenic toluene- and naphtha-degrading cultures, in addition to physiological studies, suggests that fumarate addition may be the bottleneck reaction in these three cultures. The cultures have the genetic capability of degrading structurally diverse hydrocarbons and share highly conserved and streamlined functions for anaerobic respiration and methanogenesis, unlike in situ environments impacted by hydrocarbons, which are highly variable in their functional capabilities. This observation provides future prospects for development of commercial cultures for bioremediation and biomethanization.
Language
English
DOI
doi:10.7939/R31X0G
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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