Brocadia Anammoxidans: Difference between revisions

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=Scientific Classification=
=Scientific Classification=
[[File:Anammoxdiagram.jpg|300px|thumb|right| Figure 1. Schematic drawing of anammox bacteria. Figure based on drawing from Martin Hertach.<sup>[[#References|[5]]]</sup> ]]
Domain: Bacteria
Domain: Bacteria
Phylum: Planctomycetes
Phylum: Planctomycetes
Class: Planctomycetacia
Class: Planctomycetacia
Order: Planctomycetales
Order: Planctomycetales
Family: Planctomycetaceae
Family: Planctomycetaceae
Genus: Candidatus Brocadia


Genus: ''Candidatus Brocadia''


=Introduction=
=Introduction=


Brocadia anammoxidans are anaerobic chemolithoautotrophic bacteria that belong to the order of Planctomycetes.[13] They are of special importance due to their capacity to perform the anammox (ANaerobic AMMonium OXidation) process, these bacteria were recently discovered to produce dinitrogen by removing reactive nitrogen species in marine systems.[5] Recent research being conducted in University of Nijmegen in the Netherlands has concluded that these are the only bacteria, known to date, that are able to produce hydrazine, used as rocket fuel, from waste products such as nitrite and ammonia.[10] Brocadia anammoxidans also play a leading role in removing ammonia from wastewater in the  treatment process, one of its major biotechnological applications. Residing in several different environmental habitats including: arctic sea ice, fresh water, water columns and marine sediments; these bacteria have reshaped our understanding of nitrogen cycling (figure 2) and have colossal biofuel production potential and in other various applications.[1][5][12]
''Brocadia anammoxidans'' are anaerobic chemolithoautotrophic bacteria that belong to the order of Planctomycetes.<sup>[[#References|[13]]]</sup> They are of special importance due to their capacity to perform the anammox (ANaerobic AMMonium OXidation) process, these bacteria were recently discovered to produce dinitrogen by removing reactive nitrogen species in marine systems.<sup>[[#References|[5]]]</sup> Recent research being conducted in University of Nijmegen in the Netherlands has concluded that these are the only bacteria, known to date, that are able to produce hydrazine, used as rocket fuel, from waste products such as nitrite and ammonia.[[#References|[10]]] ''Brocadia anammoxidans''  also play a leading role in removing ammonia from wastewater in the  treatment process, one of its major biotechnological applications. Residing in several different environmental habitats including: arctic sea ice, fresh water, water columns and marine sediments; these bacteria have reshaped our understanding of nitrogen cycling (figure 2) and have colossal biofuel production potential and in other various applications.<sup>[[#References|[1]]][[#References|[5]]][[#References|[12]]]</sup>
 
[[File:Figure2.jpg|300px|thumb|left| Figure 2. The nitrogen cycle. Drawing based on diagram from Jetten et al.<sup>[[#References|[3]]]</sup> ]]


=Discovery=
=Discovery=


There are three officially recognized ''Ignicoccus''  species: ''Ignicoccus hospitalis'' , '' Ignicoccus pacificus '' and '' Ignicoccus islandicus'' . The three species were initially identified by 16S rRNA gene analysis from the hydrothermal vent samples obtained from Kolbeinsey Ridge and the coast of Mexico[[#References|[1]]] . All three species have been characterized as hyperthermophiles that are also [http://en.wikipedia.org/wiki/Obligate_anaerobe obligate anaerobes] which explains the presence of ''Ignicoccus'' species near hydrothermal vents[[#References|[1]]] . None of the members of the ''Ignicoccus'' genus have been found to be [http://en.wikipedia.org/wiki/Pathogenic] pathogenic to humans.
For the past several decades, denitrification was considered to be one of the only processes responsible for the removal of ammonia from oceans.<sup>[[#References|[1]]]</sup><sup>[[#References|[10]]]</sup> In 1941, it was suggested that bacteria could use an anaerobic ammonium reaction for production and subsequent release of  nitrogen gas.<sup>[[#References|[5]]]</sup><sup>[[#References|[11]]]</sup> However, it was not until 1990 that the first direct evidence for this process was found.<sup>[[#References|[5]]]</sup> The Gist-Brocades yeast factory in Delft, Netherlands was devising a method to reduce their sulphide-rich waste and attempt to make their processes odorless.<sup>[[#References|[4]]]</sup> Sulphide levels were expected to decrease  and, according to dogma at the time, since ammonia required oxygen in order to be broken down, its concentration levels were expected to stay constant in an anaerobic reactor. The devised method worked as sulphide levels were decreasing, however, ammonia levels were surprisingly falling while nitrogen gas was being produced.<sup>[[#References|[4]]]</sup> <sup>[[#References|[6]]]</sup> Microbiologist Gijs Kuenen from Delft University of Technology was contacted and he suspected that anaerobic bacteria were combining ammonia with nitrite to form nitrogen gas and water. This marked the discovery of what his team named ''Brocadia anammoxidans'' ; anammoxidans for their unique biochemistry, and Brocadia for their place of discovery.<sup>[[#References|[4]]]</sup> <sup>[[#References|[6]]]</sup>


=Morphology=
=Cell Structure=


The members of the ''Ignicoccus''  genus are motile irregular coccoid cells that range in diameter from 1 to 3 µm. The motility observed is due to the presence of flagella, but unfortunately the polarity of the flagella is not yet fully elucidated. They are known to have an outer-membrane but no [http://en.wikipedia.org/wiki/S-layer S-layer]. This is a novel characteristic for these [http://en.wikipedia.org/wiki/Archaea Archaea] because''Ignicoccus'' are the only known Archaea that have been shown to possess an outer-membrane[[#References|[2]]] [[#References|[10]]] .
A very unique feature of ''Brocadia anammoxidans''  is the presence of an anammoxosome (figure 1), a membrane-bound compartment where the anammox reaction takes place. Findings suggested that hydrazine, a toxic and potent rocket fuel, was being produced and stored in the anammoxosome. Although very puzzling for microbiologists at first, it was later hypothesized that high energy hydrazine may be required  to drive the anammox reaction.<sup>[[#References|[2]]]</sup> Research conducted on the cell's membrane by lipid expert Jaap Sinninghe Damsté explained how these bacteria managed their toxic load without killing themselves.<sup>[[#References|[6]]]</sup> Damsté concluded that the lipids in the membrane were made from five carbon-based rings which fused together to form a "ladderane" lipid (figure 3). The high energy and instability of this structure was proposed to cause the membrane to become extremely dense, and therefore preventing hydrazine from leaking into the cell.<sup>[[#References|[2]]]</sup>


[[File:Huber-abb2.jpg|300px|thumb|left| Ultrathin section of an ''Ignicoccus hospitalis'' cell.]]
[[File:Structurediagram.jpg|300px|thumb|right|Figure 3. Chemical structure of ladderane lipid. Modified structure based on drawings from Martin Hertach. <sup>[[#References|[5]]]</sup> ]]




==Outer-Membrane==


The outer-membrane of ''Ignicoccus'' species was found to be composed of various derivatives of the typical lipid [http://en.wikipedia.org/wiki/Archaeol archaeol], including the derivative known as [http://en.wikipedia.org/wiki/Caldarchaeol caldarchaeol] [[#References|[5]]] . The outer-membrane is dominated by a pore composed of the Imp1227 protein (''Ignicoccus''  outer membrane protein 1227). The Imp1227 protein forms a large nonamer ring with a predicted pore size of 2nm[[#References|[7]]] .
=Factors affecting anammox reaction in ''Brocadia anammoxidans'' =


=Metabolism=
Some key factors affecting the activity of anaerobic ammonium oxidation include: temperature, oxygen conditions and water depth.<sup>[[#References|[5]]]</sup> ''Brocadia anammoxidans''  have an optimum temperature which ranges from 12° C to 15° C. In comparison, denitrifying bacteria's optimum temperature ranges between 25° and 30° C.<sup>[[#References|[8]]]</sup> The lower optimum temperature of ''Brocadia anammoxidans'' , allows them to out compete denitrifying bacteria in cold environments.<sup>[[#References|[5]]]</sup><sup>[[#References|[11]]]</sup>  Anammox is restricted to anoxic conditions because in the presence of oxygen, ammonium is oxidized to nitrate (nitrification). Water depth is another key factor as increasing water depth leads to a decrease in anammox rates. This is due to the fact that the anammox reaction is directly related to the presence of NO2- , which is further dependent on the mineralisation of organic matter. Since the organic matter loading of sediments decreases with increasing water depth, so does the anammox rates.<sup>[[#References|[5]]]</sup><sup>[[#References|[8]]]</sup>


''Ignicoccus'' species are [http://en.wikipedia.org/wiki/Chemolithoautotroph chemolithoautotrophs] that use molecular hydrogen as the inorganic electron donor and elemental sulphur as the inorganic terminal electron acceptor[[#References|[1]]] . The reduction of the elemental sulphur results in the production of hydrogen sulphide gas.


''Ignicoccus'' are autotrophs in that they fix their own carbon dioxide into organic molecules. The carbon dioxide fixation process they use is a novel process called [http://www.pnas.org/content/105/22/7851.full a dicarboxylate/4-hydroxybutyrate autotrophic carbon assimilation cycle] that involves 14 different enzymes[[#References|[8]]] .
=Applications=


Members of the ''Ignicoccus''  genus are able to use ammonium as a nitrogen source.
The most prevalent application of ''Brocadia anammoxidans''  lies in wastewater treatment, sewage plants, petroleum refineries and fertilizer manufacturing. These bacteria are used in breaking down the millions of litres of ammonia-rich waste that is produced. The conventional method of breaking down this ammonia-rich waste included converting ammonia into nitrite and nitrate using nitrifying bacteria, and then using denitrifying bacteria to yield nitrogen gas. The anammox reaction carried out by ''Brocadia anammoxidans''  provides huge advantages in comparison with this conventional method. The denitrifying bacteria in the conventional method require an energy source such as methanol, whereas anammox bacteria use ammonia as their fuel - and therefore there is no need for an energy source. Unlike the conventional method, anammox bacteria do not require oxygen, saving electricity that was used by machines to provide adequate oxygen levels in the conventional method. Furthermore, instead of producing CO2, anammox bacteria consume it - an environmentally friendly process.<sup>[[#References|[5]]]</sup><sup>[[#References|[6]]]</sup> Overall, this leads to an astonishing reduction of 90% in operational costs compared to the conventional method.<sup>[[#References|[3]]]</sup>
 
==Growth Conditions==
 
Because members of the ''Ignicoccus''  genus are [http://en.wikipedia.org/wiki/Hyperthermophile hyperthermophiles] and obligate anaerobes, it is not surprising that their growth conditions are very complex. They are grown in a liquid medium known as ½ SME ''Ignicoccus''  which is a solution of synthetic sea water which is then made anaerobic.
 
Grown in this media at their optimal growth temperature of 90C, the members of the ''Ignicoccus''  genus typically reach a cell density of ~4x107cells/mL[[#References|[1]]] .  
 
The addition of [http://en.wikipedia.org/wiki/Yeast_extract yeast extract] to the ½ SME media has been shown to stimulate the growth and increase maximum cell density achieved. The mechanism by which this is achieved is not known[[#References|[1]]] .
 
 
=Symbiosis=
 
''Ignicoccus hospitalis''  is the only member of the genus ''Ignicoccus'' that has been shown to have an extensive [http://en.wikipedia.org/wiki/Symbiosis symbiotic relationship] with another organism.
 
''Ignicoccus hospitalis''  has been shown to engage in symbiosis with ''Nanoarchaeum equitans'' . ''Nanoarchaeum equitans''  is a very small coccoid species with a cell diameter of 0.4 µm[[#References|[9]]] . Genome analysis has provided much of the known information about this species.
 
To further complicate the symbiotic relationship between both species, it’s been observed that the presence of ''Nanoarchaeum equitans'' on the surface of ''Ignicoccus hospitalis''  somehow inhibits the cell replication of ''Ignicoccus hospitalis'' . How or why this occurs has not yet been elucidated[[#References|[3]]] .
 
[[File:Urzwerg.jpg|300px|thumb|right| ''Ignicoccus hospitalis'' with two attached  ''Nanoarchaeum equitans'' cells.]]
 
[[File:IhNeRelationship2 jpeg.jpg|250px|thumb|left| Epifluoroscence micrographs of an ''Ignicoccus hospitalis''and ''Nanoarchaeum equitans'' coculture stained with BacLight at various time points. Living cells stain green while dead cells stain red. (A) Exponential growth phase 3.25 hours after inoculation. (B) Transition into the stationary phase 7.5 hours after inoculation. (C) Stationary phase 10 hours after inoculation. (D) Stationary phase 23 hours after inoculation.]]
 
 
==''Nanoarchaeum equitans''==
 
''Nanoarchaeum equitans'' has the smallest non-viral genome ever sequenced at 491kb[[#References|[9]]] . Analysis of the genome sequence indicates that 95% of the predicted proteins and stable RNA molecules are somehow involved in repair and replication of the cell and its genome[[#References|[3]]] .
 
Analysis of the genome also showed that ''Nanoarchaeum equitans''  lacks nearly all genes known to be required in amino acid, nucleotide, cofactor and lipid metabolism. This is partially supported by the evidence that ''Nanoarchaeum equitans''  has been shown to derive its cell membrane from its host ''Ignicoccus hospitalis''  cell membrane. The direct contact observed between ''Nanoarchaeum equitans'' and ''Ignicoccus hospitalis''  is hypothesized to form a pore between the two organisms in order to exchange metabolites or substrates (likely from ''Ignicoccus hospitalis''  towards ''Nanoarchaeum equitans'' due to the parasitic relationship). The exchange of periplasmic vesicles is not thought to be involved in metabolite or substrate exchange despite the presence of vesicles in the periplasm of ''Ignicoccus hospitalis'' .
 
These analyses of the ''Nanoarchaeum equitans'' genome support the fact of the extensive symbiotic relationship between ''Nanoarchaeum equitans'' and ''Ignicoccus hospitalis''. However, it has not yet been proven that it is a strictly parasitic relationship and further research may prove that there is a commensal relationship between the two species.


   
   
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(11) Strous, M., Van Gerven, E., Kuenen, J. G. & Jetten, M. Effects of aerobic and microaerobic conditions on anaerobic ammonium-oxidizing (Anammox) sludge. Appl. Environ. Microbiol. 63: 2446-2448 (1997).
(11) Strous, M., Van Gerven, E., Kuenen, J. G. & Jetten, M. Effects of aerobic and microaerobic conditions on anaerobic ammonium-oxidizing (Anammox) sludge. Appl. Environ. Microbiol. 63: 2446-2448 (1997).
(12) Trimmer, M., Nicholls, J.C., and Deflandre, B. (2003) Anaerobic ammonium oxidation measured in sediments along the Thames estuary, United Kingdom. Applied and Environmental Microbiology 69: 6447-6454.
(13) Van Niftrik, L.A., Fuerst, J.A., Damste, J.S.S., Kuenen, J.G., Jetten, M.S.M., Strous, M., and Ik (2004) The anammoxosome: an intracytoplasmic compartment in anammox bacteria. Fems Microbiology Letters 233: 7-13.

Latest revision as of 03:34, 15 December 2012

This student page has not been curated.

Scientific Classification

Figure 1. Schematic drawing of anammox bacteria. Figure based on drawing from Martin Hertach.[5]

Domain: Bacteria

Phylum: Planctomycetes

Class: Planctomycetacia

Order: Planctomycetales

Family: Planctomycetaceae

Genus: Candidatus Brocadia

Introduction

Brocadia anammoxidans are anaerobic chemolithoautotrophic bacteria that belong to the order of Planctomycetes.[13] They are of special importance due to their capacity to perform the anammox (ANaerobic AMMonium OXidation) process, these bacteria were recently discovered to produce dinitrogen by removing reactive nitrogen species in marine systems.[5] Recent research being conducted in University of Nijmegen in the Netherlands has concluded that these are the only bacteria, known to date, that are able to produce hydrazine, used as rocket fuel, from waste products such as nitrite and ammonia.[10] Brocadia anammoxidans also play a leading role in removing ammonia from wastewater in the treatment process, one of its major biotechnological applications. Residing in several different environmental habitats including: arctic sea ice, fresh water, water columns and marine sediments; these bacteria have reshaped our understanding of nitrogen cycling (figure 2) and have colossal biofuel production potential and in other various applications.[1][5][12]

Figure 2. The nitrogen cycle. Drawing based on diagram from Jetten et al.[3]

Discovery

For the past several decades, denitrification was considered to be one of the only processes responsible for the removal of ammonia from oceans.[1][10] In 1941, it was suggested that bacteria could use an anaerobic ammonium reaction for production and subsequent release of nitrogen gas.[5][11] However, it was not until 1990 that the first direct evidence for this process was found.[5] The Gist-Brocades yeast factory in Delft, Netherlands was devising a method to reduce their sulphide-rich waste and attempt to make their processes odorless.[4] Sulphide levels were expected to decrease and, according to dogma at the time, since ammonia required oxygen in order to be broken down, its concentration levels were expected to stay constant in an anaerobic reactor. The devised method worked as sulphide levels were decreasing, however, ammonia levels were surprisingly falling while nitrogen gas was being produced.[4] [6] Microbiologist Gijs Kuenen from Delft University of Technology was contacted and he suspected that anaerobic bacteria were combining ammonia with nitrite to form nitrogen gas and water. This marked the discovery of what his team named Brocadia anammoxidans ; anammoxidans for their unique biochemistry, and Brocadia for their place of discovery.[4] [6]

Cell Structure

A very unique feature of Brocadia anammoxidans is the presence of an anammoxosome (figure 1), a membrane-bound compartment where the anammox reaction takes place. Findings suggested that hydrazine, a toxic and potent rocket fuel, was being produced and stored in the anammoxosome. Although very puzzling for microbiologists at first, it was later hypothesized that high energy hydrazine may be required to drive the anammox reaction.[2] Research conducted on the cell's membrane by lipid expert Jaap Sinninghe Damsté explained how these bacteria managed their toxic load without killing themselves.[6] Damsté concluded that the lipids in the membrane were made from five carbon-based rings which fused together to form a "ladderane" lipid (figure 3). The high energy and instability of this structure was proposed to cause the membrane to become extremely dense, and therefore preventing hydrazine from leaking into the cell.[2]

Figure 3. Chemical structure of ladderane lipid. Modified structure based on drawings from Martin Hertach. [5]


Factors affecting anammox reaction in Brocadia anammoxidans

Some key factors affecting the activity of anaerobic ammonium oxidation include: temperature, oxygen conditions and water depth.[5] Brocadia anammoxidans have an optimum temperature which ranges from 12° C to 15° C. In comparison, denitrifying bacteria's optimum temperature ranges between 25° and 30° C.[8] The lower optimum temperature of Brocadia anammoxidans , allows them to out compete denitrifying bacteria in cold environments.[5][11] Anammox is restricted to anoxic conditions because in the presence of oxygen, ammonium is oxidized to nitrate (nitrification). Water depth is another key factor as increasing water depth leads to a decrease in anammox rates. This is due to the fact that the anammox reaction is directly related to the presence of NO2- , which is further dependent on the mineralisation of organic matter. Since the organic matter loading of sediments decreases with increasing water depth, so does the anammox rates.[5][8]


Applications

The most prevalent application of Brocadia anammoxidans lies in wastewater treatment, sewage plants, petroleum refineries and fertilizer manufacturing. These bacteria are used in breaking down the millions of litres of ammonia-rich waste that is produced. The conventional method of breaking down this ammonia-rich waste included converting ammonia into nitrite and nitrate using nitrifying bacteria, and then using denitrifying bacteria to yield nitrogen gas. The anammox reaction carried out by Brocadia anammoxidans provides huge advantages in comparison with this conventional method. The denitrifying bacteria in the conventional method require an energy source such as methanol, whereas anammox bacteria use ammonia as their fuel - and therefore there is no need for an energy source. Unlike the conventional method, anammox bacteria do not require oxygen, saving electricity that was used by machines to provide adequate oxygen levels in the conventional method. Furthermore, instead of producing CO2, anammox bacteria consume it - an environmentally friendly process.[5][6] Overall, this leads to an astonishing reduction of 90% in operational costs compared to the conventional method.[3]




References

(1) Brandes, J.A., Devol, A.H., and Deutsch, C. (2007) New developments in the marine nitrogen cycle. Chemical Reviews 107: 577-589.

(2) Elder,D.P., Snodin, D., Teasdale, A., (2001) Control and analysis of hydrazine, hydrazides and hydrazones—Genotoxic impurities in active pharmaceutical ingredients (APIs) and drug products, Journal of Pharmaceutical and Biomedical Analysis 54:900-910.

(3) Jetten, M.S.M., Cirpus, I., Kartal, B., van Niftrik, L., van de Pas-Schoonen, K.T., Sliekers, O. et al. (2005) 1994-2004: 10 years of research on the anaerobic oxidation of ammonium. Biochemical Society Transactions 33: 119-123.

(4) Kuenen, J.G., Anammox bacteria: from discovery to application. Nature Reviews Microbiology 64: 320 (2008).

(5) Martin, H., Anaerobic Ammonium Oxidation (Anammox) – A new sink in the marine nitrogen cycle. ETH Zurich, (2008).

(6) Pilcher, S., Microbiology: Pipe Dreams. Nature Publishing Group 437: 1227-1228 (2005).

(7) Rysgaard, S., Glud, R.N., Risgaard-Petersen, N., Dalsgaard, T., and Jt (2004b) Denitrification and anammox activity in Arctic marine sediments. Limnology and Oceanography 49: 1493-1502.

(8) Saad, O., Conrad, R., and Kg (1993) TEMPERATURE-DEPENDENCE OF NITRIFICATION, DENITRIFICATION, AND TURNOVER OF NITRIC-OXIDE IN DIFFERENT SOILS. Biology and Fertility of Soils 15: 21-27.

(9) Schubert, C.J., Durisch-Kaiser, E., Wehrli, B., Thamdrup, B., Lam, P., and Kuypers, M.M.M. (2006) Anaerobic ammonium oxidation in a tropical freshwater system (Lake Tanganyika). Environmental Microbiology 8: 1857-1863.

(10) Stefan,H., Aller, R., Canfield,D.E.,Dalsgaard (2005)Nitrogen removal in marine environments: recent findings and future research challenges, Marine Chemistry 94: 125-145.

(11) Strous, M., Van Gerven, E., Kuenen, J. G. & Jetten, M. Effects of aerobic and microaerobic conditions on anaerobic ammonium-oxidizing (Anammox) sludge. Appl. Environ. Microbiol. 63: 2446-2448 (1997).

(12) Trimmer, M., Nicholls, J.C., and Deflandre, B. (2003) Anaerobic ammonium oxidation measured in sediments along the Thames estuary, United Kingdom. Applied and Environmental Microbiology 69: 6447-6454.

(13) Van Niftrik, L.A., Fuerst, J.A., Damste, J.S.S., Kuenen, J.G., Jetten, M.S.M., Strous, M., and Ik (2004) The anammoxosome: an intracytoplasmic compartment in anammox bacteria. Fems Microbiology Letters 233: 7-13.