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Permanent link (DOI): https://doi.org/10.7939/R3KP7V53Q

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Denitrification and the hypoxic response in obligate aerobic methane-oxidizing bacteria Open Access

Descriptions

Other title
Subject/Keyword
denitrification
nitrous oxide
Methane
Methylomonas
Methylomicrobium
nitrite reduction
Methanotroph
nitrate respiration
hypoxia
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Kits, Kerim D
Supervisor and department
Stein, Lisa (Biological Sciences)
Examining committee member and department
Stein, Lisa (Biological Sciences)
Boucher, Yan (Biological Sciences)
Dennis, Jonathan (Biological Sciences)
Owttrim, George (Biological Sciences)
Belosevic, Mike (Biological Sciences)
Voordouw, Gerrit (Biological Scicnes)
Department
Department of Biological Sciences
Specialization
Microbiology and Biotechnology
Date accepted
2016-08-23T15:49:17Z
Graduation date
2016-06:Fall 2016
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Aerobic methanotrophic bacteria lessen the impact of the greenhouse gas methane (CH4) not only because they are a sink for atmospheric methane but also because they oxidize it before it is emitted to the atmospheric reservoir. Aerobic methanotrophs, unlike anaerobic methane oxidizing archaea, have a dual need for molecular oxygen (O2) for respiration and CH4 oxidation. Nevertheless, methanotrophs are highly abundant and active in environments that are extremely hypoxic and even anaerobic. While the O2 requirement in these organisms for CH4 oxidation is inflexible, recent genome sequencing efforts have uncovered the presence of putative denitrification genes in many aerobic methanotrophs. Being able to use two different terminal electron acceptors – hybrid respiration – would be massively advantageous to aerobic methanotrophs as it would allow them to halve their O2 requirement. But, the function of these genes that hint at an undiscovered respiratory anaerobic metabolism is unknown. Moreover, past work on pure cultures of aerobic methanotrophs ruled out the possibility that these organisms denitrify. An organism that can couple CH4 oxidation to NO3- respiration so far does not exist in pure culture. So while the role of aerobic methanotrophs in the carbon cycle is appreciated, the hypoxic metabolism and contribution of these specialized microorganisms to the nitrogen cycle is not understood. Here we demonstrate using cultivation dependent approaches, microrespirometry, and whole genome, transcriptome, and proteome analysis that an aerobic methanotroph – Methylomonas denitrificans FJG1 – couples CH4 oxidation to NO3- respiration with N2O as the terminal product via the intermediates NO2- and NO. Whole transcriptome and proteome analysis reveals that respiratory nitrate (Nar, Nap), nitrite (Nir), and nitric oxide (Nor) reductases are expressed and upregulated at the transcript and protein levels under denitrifying conditions. Physiological analysis of denitrifying cultures of M. denitrificans FJG1 also confirms that NO3- respiration is bioenergetically advantageous. We also confirm denitrification activity and upregulation of denitrification gene expression in another obligate methanotroph – Methylomicrobium album BG8 – and also show that this activity is supported by a diverse array of energy sources. Formaldehyde fermentation has been identified by others as an important adaptation of aerobic methanotrophs to hypoxia. We additionally illustrate here that M. denitrificans FJG1 can also ferment formaldehyde and that nitrate respiration and fermentation occur simultaneously during hypoxia. The work herein is a significant contribution to our knowledge of how aerobic methanotrophs affect greenhouse gas flux though CH4 oxidation in low O2 environments and through N2O production via denitrification.
Language
English
DOI
doi:10.7939/R3KP7V53Q
Rights
This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for the purpose of private, scholarly or scientific research. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.
Citation for previous publication
Kits, K. D., Klotz, M. G., and Stein, L. Y. (2015). Methane oxidation coupled to nitrate reduction under hypoxia by the Gammaproteobacterium Methylomonas denitrificans, sp. nov. type strain FJG1. Environ. Microbiol. 17, 3219–3232. doi:10.1111/1462-2920.12772.Kits, K. D., Campbell, D. J., Rosana, A. R., and Stein, L. Y. (2015). Diverse electron sources support denitrification under hypoxia in the obligate methanotroph Methylomicrobium album strain BG8. Front. Microbiol. 6, 1072. doi:10.3389/fmicb.2015.01072.

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