Methanosaeta thermophila: Difference between revisions

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Methanosaeta thermophila 
{{Curated}}
{{Biorealm Genus}}


● ~Classification~
==Classification==
Organism Name: Methanosaeta thermophila PT
Domain: Archaea
Domain: Archaea
Phylum: Euryarchaeota
                    Phylum: Euryarchaeota
Class: Methanomicrobia
                      Class: Methanomicrobia
Order: Methanosarcinales
                      Order: Methanosarcinales
Family: Methanosaetaceae
                    Family: Methanosaetaceae
Genus: Methanosaeta
                    Genus: Methanosaeta
Species: Methanothrix thermophila
                  Species:     Methanothrix thermophila
Genus Species Strain: Methanosaeta thermophila PT
      Genus Species Strain:     Methanosaeta thermophila PT
Name History: Synonyms: Methanothrix thermophila PT
              Name History: Synonyms: Methanothrix thermophila PT
                Methanothrix thermophila DSM 6194  
                                          Methanothrix thermophila DSM 6194  
Equivalent names: Methanosaeta thermophila strain PT
          Equivalent names:     Methanosaeta thermophila strain PT
                          Methanosaeta thermophila str. PT
                                Methanosaeta thermophila strain PT


==Description and Significance==
●~Description and Significance~


             Methanosaeta thermophila are a diverse group of widely distributed anaerobic archaea that occur in anaerobic environments, such as the intestinal tracts of animals, freshwater and marine sediments, sewage, anaerobic biofilms, and anaerobic sediments, and are methanogens, which means they are capable of producing methane from a limited number of substrates, including carbon dioxide and hydrogen, acetate, and methylamines. Methanosaeta thermophila are nonmotile, nonsporulating, and thermophilic, which means they thrive at temperatures of 50ºC or higher.
              
This microbe was discovered by a molecular technique using fluorogenic PCR to identify methanotrophic components and activity in marine anoxic microbial communities by identifying and quantifying the mcrA genes. Molecular analysis was performed by amplification and clone analysis of 16S rRNA and mcrA genes.  The mcrA genes (encoding the methyl coenzyme M reductase, specific to methanogenic archaea), are specific to the various phylogenetic groups of methanotropic Archaea. Methanosaeta thermophila was identified among the microbial communities in cold deep sediments and methane seepages of Omine Ridge in the Nankai Trough accretionary prism. 
The addition of Methanosaeta to the methanoarchaeal genome sequence compilation offers an opportunity to gain significant insight into this difficult microbe and the unprecedented use of comparative genomic approaches allows one to address the nature of these microbes and their biological impact and potential. Because they are methanogens, they are an important source of natural gas and have potential as a producer of biofuels.


            ''Methanosaeta thermophila'' are nonmotile, nonsporulating, and
  thermophilic, which means they thrive at temperatures of 50ºC or higher.
  The addition of ''Methanosaeta'' to the methanoarchaeal genome
  sequence compilation offered an opportunity to gain significant insight into
  this intricate microbe and the unique use of comparative genomic approaches
  allows one to address the nature of these specific microbes and their biological
  influence and capability. Because these microbes are methanogens, they serve
  an important role as the producers of natural gas and have potential as
  creators of biofuels (fuels derived from a biomass).


● ~Genome Structure~
==Genome Structure==


The Methanosaeta thermophila’s genome has been entirely sequenced.  These microbes possess circular chromosomes and do not contain plasmids.  
The ''Methanosaeta thermophila'' genome has been entirely sequenced.   
 
  These microbes possess circular chromosomes and do not contain plasmids.  
Genome Sequence: RS: NC_008553     
  (The following genome sequence information is from reference # 11)
Genome Sequence Length: 1879471  
        Genome Sequence: RS: NC_008553     
Statistics: Number of nucleotides: 1879471
        Genome Sequence Length: 1879471  
                Number of protein genes: 1696
        Statistics: Number of nucleotides: 1879471
                Number of RNA genes: 51
                    Number of protein genes: 1696
Genome Statistics
                    Number of RNA genes: 51
Number % of Total
       
DNA, total number of bases 100.00%
==Cell Structure and Metabolism==
        DNA coding number of bases
''Methanosaeta thermophila'' is circular (coccus), with one inner membrane
        DNA G+C number of bases 0.00% 1
  and one cell wall. This microbe does not interact with other organisms, grows
                       
  extremely slow, does not contain plasmids, does not possess flagella, but they
DNA scaffolds 0 100.00%
  do however produce gas vacuoles to help them move in aquatic environments. Gas
                       
  vacuoles are cavities within the cytoplasm, which contain a gas similar to that
Genes total number 0 100.00%
  of their surrounding atmosphere. These vacuoles serve as flotation devices
        Protein coding genes 0 0.00%
  because they decrease in size when subjected to increased hydrostatic pressure.  
        RNA genes 0 0.00%
  So although they are nonmotile, their gas vacuoles allow some degree of
                rRNA genes 0 0.00%
  flexibility in regards to how much movement they have in aquatic environments.  
                        5S rRNA 0 0.00%
      ''Methanosaeta thermophila'' obtain their energy as a “thermophilic
                        16S rRNA 0 0.00%
  obligately-aceticlastic methane-producing archaeon,” which means that they
                        18S rRNA 0 0.00%
  produce methane from acetate (4). Although approximately two-thirds of all
                        23S rRNA 0 0.00%
  methane is derived from the methyl group of acetate, ''Methanosaeta'' are able
                        28S rRNA 0 0.00%
  to utilize acetate as a substrate for methanogenesis. ''Methanosarcina'' is
                tRNA genes 0 0.00%
  the only other genus of methanoarchaea that are capable of utilizing acetate
                Other RNA genes 0 0.00%
  as a substrate, as well as using H2/CO2, dimethylsulfide, and and methanethiol
        Genes with function prediction 0 0.00%
  compounds as substratesUnlike the faster-growing ''Methanosarcina'', which
        Genes without function prediction 0 0.00%
  prefers methylated compounds to acetate, ''Methanosaeta'' is a slow-growing
  specialist that utilizes acetate only. 
                Genes w/o function with similarity 0 0.00%
                Genes w/o function w/o similarity 0 0.00%
        Pseudo Genes 0 0.00%
        Genes assigned to enzymes 0 0.00%
        Genes connected to KEGG pathways 0 0.00%
        Genes not connected to KEGG pathways 0 0.00%
        Genes in ortholog clusters 0 0.00%
        Genes in paralog clusters 0 0.00%
        Genes in COGs 0 0.00%
        Genes in Pfam 0 0.00%
        Genes in TIGRfam 0 0.00%
        Genes in InterPro 0 0.00%
        Genes with IMG Terms 0 0.00%
        Genes in IMG Pathways 0 0.00%
        Obsolete Genes 0 0.00%
        Revised Genes 0 0.00%
Pfam clusters 0.00%
Paralogous groups 100.00%
Orthologous groups 0.00%
Map of chromosome
 
  Map of Methanosaeta thermophila PT: NC_008553
Legend: From outside to the center
Genes on forward strand (color by COG categories)
Genes on reverse strand (color by COG categories)
RNA genes (tRNAs green, sRNAs red)
GC content
GC skew


COG Coloring Selection
        
Color code of function category for top COG is shown below.
COG Code COG Function Definition
[A] RNA processing and modification
[B] Chromatin structure and dynamics
[C] Energy production and conversion
[D] Cell cycle control, cell division, chromosome partitioning
[E] Amino acid transport and metabolism
[F] Nucleotide transport and metabolism
[G] Carbohydrate transport and metabolism
[H] Coenzyme transport and metabolism
[I] Lipid transport and metabolism
[J] Translation, ribosomal structure and biogenesis
[K] Transcription
[L] Replication, recombination and repair
[M] Cell wall/membrane/envelope biogenesis
[N] Cell motility
[O] Posttranslational modification, protein turnover, chaperones
[P] Inorganic ion transport and metabolism
[Q] Secondary metabolites biosynthesis, transport and catabolism
[R] General function prediction only
[S] Function unknown
[T] Signal transduction mechanisms
[U] Intracellular trafficking, secretion, and vesicular transport
[V] Defense mechanisms
[W] Extracellular structures
[Y] Nuclear structure
[Z] Cytoskeleton
NA Not assigned
 
 
● ~Cell Structure and Metabolism~ 
 
Methanosaeta thermophila is circular (coccus), with one inner membrane and one cell wall.  This microbe does not interact with other organisms, grows extremely slow, does not contain plasmids, does not possess flagella, but they do however produce gas vacuoles to help them move in aquatic environments. Gas vacuoles are cavities within the cytoplasm, which contain gas similar to that of their surrounding atmosphere. These vacuoles serve as floating devices because they decrease in size when subjected to increased hydrostatic pressure.        
Methanosaeta thermophila obtain their energy production as a thermophilic obligately-aceticlastic methane-producing archaeon, which means that they produce methane from acetate.  Although approximately two-thirds of all biogenic methane is derived from the methyl group of acetate, Methanosaeta is one of only two genera of the methanoarchaea able to utilize acetate as a substrate for methanogenesis.  Methanosarcina, the only other genus of methanoarchaea able to utilize acetate, can use H2/CO2, dimethylsulfide, and methanethiol as well as methylated C1 compounds as substrates.  Unlike the faster-growing Methanosarcina, which prefers methylated compounds to acetate, Methanosaeta is a specialist that utilizes only acetate. 
       
● ~Ecology~ 


The environment at which Methanosaeta thermophiles are found is aquatic and they exhibit optimal growth between 55-60°C. Although they are present in many environments, such as anaerobic digesters, anaerobic biofilms, sediments, and anaerobic sludges, they are predominantly found in rice paddies. Acetate is the most important substrate for methanogenesis in rice paddies and studies have shown that the concentration of acetate in flooded rice paddies is in the 5-100 mM range, and are the predominant acetate-utilizing methanoarchaea in flooded rice paddies. 
Methanosaeta species also dominate the methanogenic population of microbial consortia in granular sludge digestors, codigestors treating municipal solid waste and sewage sludge, upflow anaerobic sludge blanket reactors, and anaerobic baffled reactors.  During start-up of anaerobic bioreactors, Methanosarcina species are often prevalent due to the high acetate concentration; however, as bioreactors stabilize and achieve optimum performance the acetate concentration decreases and Methanosaeta species replace Methanosarcina .


==Ecology== 


        The environment at which ''Methanosaeta thermophiles'' are found is
  aquatic (living and growing in water) and they exhibit optimal growth between
  55-60°C.
  ''Methanosaeta species are the most prevalent methanogenic archaea of
  the microbial population in numerous environments, including rough sludge
  digesters, solid wastes, sewage slush,and anaerobic reactors.  During activation
  of anaerobic bioreactors, ''Methanosaeta'' species are widespread due to the
  high acetate concentration. However, as bioreactors become stable and attain
  their peak performance, the acetate concentration decreases, as well as the
  ''Methanosaeta'' population.


● ~Pathology~
===Pathology===


Methanosaeta thermofiles are not pathogens and therefore are not disease causing microbes. To this date, there are no known archaea that are pathogens.  
        ''Methanosaeta thermophila''are not pathogens and therefore are not disease  
  causing microbes. To this date, there are no known archaea that are pathogens.


● ~Application to Biotechnology~ 
==Current Research==


Methanosaeta thermofiles are relevant for biotechnology, energy production, and as major producers as biofuels. M. thermophila is a source of natural gas as methanogens could be useful producers of biofuels because they can metabolize acetate into methane by reducing the methyl group to CH4 with electrons derived from oxidation of the carbonyl group to CO2. With biofuels, fuel is produced from renewable resources, such as plant biomass, vegetable oils, and treated municipal and industrial wastes. The use of biofuels as an additive to petroleum-based fuels can result in cleaner burning with less emission of carbon monoxide and particulates.  
        Although ''Methanosaeta'' continues to be comprehensively studied both
Methanosaeta thermofiles are also contributors to biogas; a mixture of methane and carbon dioxide produced by the anaerobic decomposition of organic matter such as sewage and municipal wastes by bacteria. This is important because it is used in the generation of hot water and electricity.
  biochemically and genetically, studies have decreased due to its slow growth
  (up to 12 days doubling time) and lower growth yield than other microbes. 
  One current study of this microbe by Alber, B.E. and Ferry J.G., determined
  that ''Methanosaeta thermophila'' archaea produce a carbonic anhydrase, which
  is an enzyme that catalyzes the reaction of water with carbon dioxide. The
  carbonic anhydrase(abbreviated CA) from acetate-grown ''Methanosaeta
  thermophila'' was purified. The molecular mass of the enzyme (CA) was
  determined via gel filtration chromatography. The results indicated that
  this particular CA represented a distinct class of CA’s and provided a
  foundation to determine the unique roles for CA in acetotrophic anaerobes.
        Another study detected central glutamates in the acetate kinase from  
  the ''Methanosaeta thermophila''. Acetate kinase is an enzyme which catalyzes
  the reversible phosphorylation (adding a phosphate group) of acetate. The
  suggested mechanism denoted an unspecified glutamate residue was phosphorylated,  
  and the “alignment of the amino acid sequences for the acetate kinases from  
  ''E. coli'' (Bacteria domain), ''Methanosarcina thermophila'' (Archaea domain),  
  and four other phylogenetically divergent microbes revealed high identity
  which included five glutamates,” (9). These glutamates were substituted in
  the ''M. thermophila'' enzyme to determine if they were required for catalysis.  
  The substituted enzymes were tagged and created in ''E. coli'' and purified by  
  metal affinity chromatography. The substituted enzymes produced undetectable
  kinase activity.  These results imply that the glutamates in the acetate
  kinase were in fact required for catalysis, which supports the original
  suggested mechanism.


● ~Current Research~
==References==


Although Methanosarcina continues to be extensively studied both biochemically and genetically, studies on Methanosaeta have lapsed due to its slow growth (2-12 day doubling time) and lower growth yield. One current study of this microbe by Alber, BE and Ferry JG, determined that Methanosaeta thermofile produce a carbonic anhydrase. The carbonic anhydrase (CA) from acetate-grown Methanosarcina thermophila was purified. The molecular mass of the enzyme was determined via gel filtration chromatography. The results indicate that this archaeal CA represents a distinct class of CAs and provide a basis to determine physiological roles for CA in acetotrophic anaerobes.
  1) Copeland A., Lucas S., Lapidus A., Barry K., Detter J.C., Glavina del Rio T.,
Another study identified essential glutamates in the acetate kinase from the Methanosaeta thermophila. Acetate kinase catalyzes the reversible phosphorylation of acetate. The proposed mechanism indicated an unspecified glutamate residue was phosphorylated. Alignment of the amino acid sequences for the acetate kinases from E. coli (Bacteria domain), Methanosarcina thermophila (Archaea domain), and four other phylogenetically divergent microbes revealed high identity which included five glutamates. These glutamates were replaced in the M. thermophila enzyme to determine if any were essential for catalysis. The altered enzymes were tagged and produced in E. coli and purified by metal affinity chromatography. The enzymes showed either undetectable or extremely low kinase activity, suggesting that the glutamates in the acetate kinase were essential for catalysis, which supports the proposed mechanism.
    Hammon N., Israni S., Pitluck S., Chain P., Malfatti S., Shin M., Vergez L.,
A third study titled, “A genetic system for Archaea of the genus Methanosarcina: Liposome-mediated transformation and construction of shuttle vectors,” explored new methods that allow genetic analysis in Archaea. Several autonomously replicating plasmid shuttle vectors were constructed. These vectors replicate in 9 of 11 Methanosarcina strains tested and in Escherichia coli. A transformation system was established where DNA was introduced by liposomes, which allowed for extremely efficient transformation frequencies During the course of this work, the complete DNA sequence was determined.
    Schmutz J.,  Larimer F., Land M., Hauser L., Kyrpides N., Kim E., Smith K.S.,
    Ingram-Smith C., Richardson P.; "Complete sequence of Methanosaeta thermophila
    PT."; Submitted (OCT-2006) to the EMBL/GenBank/DDBJ databases.
  2) Identification of Essential Glutamates in the Acetate Kinase from
    Methanosarcina thermophila. Singh-Wissmann K, Ingram-Smith C, Miles RD, Ferry
    JG. J Bacteriol. 1998 Mar; 180(5): 1129-1134.
  3) Carbonic anhydrase is an ancient enzyme widespread in prokaryotes. Smith KS,
    Jakubzick C, Whittam TS, Ferry JG. Proc Natl Acad Sci U S A. 1999 Dec 21; 96(26):
    15184-15189.
  4) A carbonic anhydrase from the archaeon Methanosarcina thermophila.
    Alber BE, Ferry JG. Proc Natl Acad Sci U S A. 1994 Jul 19; 91(15): 6909-6913.
  6) NCBI, Joint Genome Institute, Unpublished, October 25, 2006, Richardson P
      http://www.genomesonline.org/DBs/goldtable.txt
  7) http://www.genome.jp/kegg-bin/show_organism?org=mtp
  8) ftp://ftp.ncbi.nih.gov/genomes/Bacteria/Methanosaeta_thermophila_PT/
  9) http://genome.jgi-psf.org/draft_microbes/metth/metth.info.html
10) NCBI/RefSeq:NC_008553 http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val
    =NC_008553
11) Map of Chromosome: http://img.jgi.doe.gov/cgi-bin/pub/main.cgi?section=TaxonCircMaps&page
    =circMaps&taxon_oid=639633038&pidt=25318.1178126134
12) http://genome.jgi-psf.org/finished_microbes/metth/metth.home.html
13) http://genome.jgi-psf.org/finished_microbes/metth/metth.home.html
14) http://expasy.org/sprot/hamap/METTP.html
15) http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=349307


● ~References~


1) Copeland A., Lucas S., Lapidus A., Barry K., Detter J.C., Glavina del Rio T., Hammon N., Israni S., Pitluck S., Chain P., Malfatti S., Shin M., Vergez L., Schmutz J., Larimer F., Land M., Hauser L., Kyrpides N., Kim E., Smith K.S., Ingram-Smith C., Richardson P.; "Complete sequence of Methanosaeta thermophila PT."; Submitted (OCT-2006) to the EMBL/GenBank/DDBJ databases.
KMG
2) Identification of Essential Glutamates in the Acetate Kinase from Methanosarcina thermophila. Singh-Wissmann K, Ingram-Smith C, Miles RD, Ferry JG. J Bacteriol. 1998 Mar; 180(5): 1129-1134.
3) Carbonic anhydrase is an ancient enzyme widespread in prokaryotes. Smith KS, Jakubzick C, Whittam TS, Ferry JG. Proc Natl Acad Sci U S A. 1999 Dec 21; 96(26): 15184-15189.
4) A carbonic anhydrase from the archaeon Methanosarcina thermophila.
Alber BE, Ferry JG. Proc Natl Acad Sci U S A. 1994 Jul 19; 91(15): 6909-6913.
5) A genetic system for Archaea of the genus Methanosarcina: Liposome-mediated transformation and construction of shuttle vectors. Metcalf WW, Zhang JK, Apolinario E, Sowers KR, Wolfe RS. Proc Natl Acad Sci U S A. 1997 Mar 18; 94(6): 2626-2631.
PMCID: 20139
1)  NCBI, Joint Genome Institute, Unpublished, October 25, 2006, Richardson P http://www.genomesonline.org/DBs/goldtable.txt
2) http://www.genome.jp/kegg-bin/show_organism?org=mtp
3) ftp://ftp.ncbi.nih.gov/genomes/Bacteria/Methanosaeta_thermophila_PT/
4) http://genome.jgi-psf.org/draft_microbes/metth/metth.info.html
5) NCBI/RefSeq:NC_008553
http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=NC_008553
6) Map of Chromosome: http://img.jgi.doe.gov/cgi-bin/pub/main.cgi?section=TaxonCircMaps&page=circMaps&taxon_oid=639633038&pidt=25318.1178126134
7) http://genome.jgi-psf.org/finished_microbes/metth/metth.home.html
8) http://genome.jgi-psf.org/finished_microbes/metth/metth.home.html
9) http://expasy.org/sprot/hamap/METTP.html
10) http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=349307

Latest revision as of 15:25, 7 July 2011

This is a curated page. Report corrections to Microbewiki.

A Microbial Biorealm page on the genus Methanosaeta thermophila

Classification

Domain: Archaea

                   Phylum:	Euryarchaeota
                     Class:	Methanomicrobia
                     Order:	Methanosarcinales
                   Family:	Methanosaetaceae
                    Genus: 	Methanosaeta
                  Species:     Methanothrix thermophila
     Genus Species Strain:     Methanosaeta thermophila PT
             Name History:	Synonyms: Methanothrix thermophila PT
                                         Methanothrix thermophila DSM 6194 
         Equivalent names:     Methanosaeta thermophila strain PT
                               Methanosaeta thermophila strain PT

Description and Significance

           Methanosaeta thermophila are nonmotile, nonsporulating, and 
 thermophilic, which means they thrive at temperatures of 50ºC or higher.
 The addition of Methanosaeta to the methanoarchaeal genome 
 sequence compilation offered an opportunity to gain significant insight into 
 this intricate microbe and the unique use of comparative genomic approaches 
 allows one to address the nature of these specific microbes and their biological 
 influence and capability. Because these microbes are methanogens, they serve 
 an important role as the producers of natural gas and have potential as 
 creators of biofuels (fuels derived from a biomass).

Genome Structure

The Methanosaeta thermophila genome has been entirely sequenced.

 These microbes possess circular chromosomes and do not contain plasmids. 
 (The following genome sequence information is from reference # 11)
        Genome Sequence: RS: NC_008553     
        Genome Sequence Length: 1879471 
        Statistics: Number of nucleotides: 1879471
                    Number of protein genes: 1696
                    Number of RNA genes: 51
        

Cell Structure and Metabolism

Methanosaeta thermophila is circular (coccus), with one inner membrane
 and one cell wall. This microbe does not interact with other organisms, grows 
 extremely slow, does not contain plasmids, does not possess flagella, but they 
 do however produce gas vacuoles to help them move in aquatic environments. Gas
 vacuoles are cavities within the cytoplasm, which contain a gas similar to that 
 of their surrounding atmosphere. These vacuoles serve as flotation devices 
 because they decrease in size when subjected to increased hydrostatic pressure. 
 So although they are nonmotile, their gas vacuoles allow some degree of 
 flexibility in regards to how much movement they have in aquatic environments.    
      Methanosaeta thermophila obtain their energy as a “thermophilic 
 obligately-aceticlastic methane-producing archaeon,” which means that they 
 produce methane from acetate (4).  Although approximately two-thirds of all 
 methane is derived from the methyl group of acetate, Methanosaeta are able
 to utilize acetate as a substrate for methanogenesis. Methanosarcina is 
 the only other genus of methanoarchaea that are capable of utilizing acetate
 as a substrate, as well as using H2/CO2, dimethylsulfide, and and methanethiol 
 compounds as substrates.  Unlike the faster-growing Methanosarcina, which 
 prefers methylated compounds to acetate, Methanosaeta is a slow-growing 
 specialist that utilizes acetate only.  



Ecology

       The environment at which Methanosaeta thermophiles are found is 
 aquatic (living and growing in water) and they exhibit optimal growth between
 55-60°C. 
 Methanosaeta species are the most prevalent methanogenic archaea of 
 the microbial population in numerous environments, including rough sludge 
 digesters, solid wastes, sewage slush,and anaerobic reactors.  During activation 
 of anaerobic bioreactors, Methanosaeta species are widespread due to the 
 high acetate concentration. However, as bioreactors become stable and attain 
 their peak performance, the acetate concentration decreases, as well as the 
 Methanosaeta population.

Pathology

       Methanosaeta thermophilaare not pathogens and therefore are not disease 
 causing microbes. To this date, there are no known archaea that are pathogens.

Current Research

        Although Methanosaeta continues to be comprehensively studied both
 biochemically and genetically, studies have decreased due to its slow growth
 (up to 12 days doubling time) and lower growth yield than other microbes.  
 One current study of this microbe by Alber, B.E. and Ferry J.G., determined 
 that Methanosaeta thermophila archaea produce a carbonic anhydrase, which 
 is an enzyme that catalyzes the reaction of water with carbon dioxide. The 
 carbonic anhydrase(abbreviated CA) from acetate-grown Methanosaeta 
 thermophila was purified. The molecular mass of the enzyme (CA) was 
 determined via gel filtration chromatography. The results indicated that 
 this particular CA represented a distinct class of CA’s and provided a 
 foundation to determine the unique roles for CA in acetotrophic anaerobes.
        Another study detected central glutamates in the acetate kinase from 
 the Methanosaeta thermophila. Acetate kinase is an enzyme which catalyzes
 the reversible phosphorylation (adding a phosphate group) of acetate. The
 suggested mechanism denoted an unspecified glutamate residue was phosphorylated, 
 and the “alignment of the amino acid sequences for the acetate kinases from 
 E. coli (Bacteria domain), Methanosarcina thermophila (Archaea domain), 
 and four other phylogenetically divergent microbes revealed high identity 
 which included five glutamates,” (9). These glutamates were substituted in
 the M. thermophila enzyme to determine if they were required for catalysis. 
 The substituted enzymes were tagged and created in E. coli and purified by 
 metal affinity chromatography. The substituted enzymes produced undetectable 
 kinase activity.  These results imply that the glutamates in the acetate
 kinase were in fact required for catalysis, which supports the original 
 suggested mechanism.

References

 1) Copeland A., Lucas S., Lapidus A., Barry K., Detter J.C., Glavina del Rio T.,
    Hammon N., Israni S., Pitluck S., Chain P., Malfatti S., Shin M., Vergez L.,
    Schmutz J.,  Larimer F., Land M., Hauser L., Kyrpides N., Kim E., Smith K.S., 
    Ingram-Smith C., Richardson P.; "Complete sequence of Methanosaeta thermophila 
    PT."; Submitted (OCT-2006) to the EMBL/GenBank/DDBJ databases.
 2) Identification of Essential Glutamates in the Acetate Kinase from 
    Methanosarcina thermophila. Singh-Wissmann K, Ingram-Smith C, Miles RD, Ferry
    JG. J Bacteriol. 1998 Mar; 180(5): 1129-1134.
 3) Carbonic anhydrase is an ancient enzyme widespread in prokaryotes. Smith KS,
    Jakubzick C, Whittam TS, Ferry JG. Proc Natl Acad Sci U S A. 1999 Dec 21; 96(26): 
    15184-15189.
 4) A carbonic anhydrase from the archaeon Methanosarcina thermophila.
    Alber BE, Ferry JG. Proc Natl Acad Sci U S A. 1994 Jul 19; 91(15): 6909-6913.
 6)  NCBI, Joint Genome Institute, Unpublished, October 25, 2006, Richardson P 
     http://www.genomesonline.org/DBs/goldtable.txt
 7) http://www.genome.jp/kegg-bin/show_organism?org=mtp
 8) ftp://ftp.ncbi.nih.gov/genomes/Bacteria/Methanosaeta_thermophila_PT/
 9) http://genome.jgi-psf.org/draft_microbes/metth/metth.info.html
10) NCBI/RefSeq:NC_008553 http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val
    =NC_008553
11) Map of Chromosome: http://img.jgi.doe.gov/cgi-bin/pub/main.cgi?section=TaxonCircMaps&page
    =circMaps&taxon_oid=639633038&pidt=25318.1178126134
12) http://genome.jgi-psf.org/finished_microbes/metth/metth.home.html
13) http://genome.jgi-psf.org/finished_microbes/metth/metth.home.html 
14) http://expasy.org/sprot/hamap/METTP.html
15) http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=349307


KMG