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Literature summary for 1.5.98.1 extracted from

  • Shima, S.; Thauer, R.K.
    Tetrahydromethanopterin-specific enzymes from Methanopyrus kandleri (2001), Methods Enzymol., 331, 317-353.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information relatively high concentrations of lyotropic salt is required for activity, 200fold activation Methanopyrus kandleri

Cloned(Commentary)

Cloned (Comment) Organism
gene mtd, DNA and amino acid sequence determination and analysis, functional overexpression in Escherichia coli strain BL21(DE3) Methanopyrus kandleri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.006
-
methylenetetrahydromethanopterin
-
Methanosarcina barkeri
0.013
-
oxidized coenzyme F420
-
Archaeoglobus fulgidus
0.017
-
methylenetetrahydromethanopterin
-
Archaeoglobus fulgidus
0.02
-
oxidized coenzyme F420 native enzyme, pH 6.0, 65°C, N2-atmosphere Methanopyrus kandleri
0.025
-
oxidized coenzyme F420
-
Methanosarcina barkeri
0.033
-
methylenetetrahydromethanopterin
-
Methanothermobacter thermautotrophicus
0.065
-
oxidized coenzyme F420
-
Methanothermobacter thermautotrophicus
0.08
-
methylenetetrahydromethanopterin native enzyme, pH 6.0, 65°C, N2-atmosphere Methanopyrus kandleri

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29644
-
4 * 32000, approximately, SDS-PAGE, 4 * 29644, sequence calculation Archaeoglobus fulgidus
29644
-
6 * 36000, approximately, SDS-PAGE, 6 * 29644, sequence calculation Methanothermobacter thermautotrophicus
30305
-
x * 30305, sequence calculation Methanocaldococcus jannaschii
31000
-
8 * 31000, approximately, SDS-PAGE Methanosarcina barkeri
31383
-
8 * 36000, approximately, SDS-PAGE, 8 * 31383, sequence calculation Methanopyrus kandleri
32000
-
4 * 32000, approximately, SDS-PAGE, 4 * 29644, sequence calculation Archaeoglobus fulgidus
36000
-
6 * 36000, approximately, SDS-PAGE, 6 * 29644, sequence calculation Methanothermobacter thermautotrophicus
36000
-
8 * 36000, approximately, SDS-PAGE, 8 * 31383, sequence calculation Methanopyrus kandleri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Methanosarcina barkeri enzyme is involved in CO2 reduction to CH4 ?
-
?
additional information Methanothermobacter thermautotrophicus enzyme is involved in CO2 reduction to CH4 ?
-
?
additional information Methanocaldococcus jannaschii enzyme is involved in CO2 reduction to CH4 ?
-
?
additional information Archaeoglobus fulgidus enzyme is involved in CO2 reduction to CH4 ?
-
?
additional information Methanopyrus kandleri enzyme is involved in CO2 reduction to CH4 ?
-
?

Organism

Organism UniProt Comment Textmining
Archaeoglobus fulgidus
-
-
-
Methanocaldococcus jannaschii
-
-
-
Methanopyrus kandleri
-
hyperthermophilic archeon, methanogen, gene mtd
-
Methanosarcina barkeri
-
-
-
Methanothermobacter thermautotrophicus
-
-
-

Oxidation Stability

Oxidation Stability Organism
O2 causes slow inactivation Methanopyrus kandleri

Purification (Commentary)

Purification (Comment) Organism
native enzyme, multistep procedure involving ammonium sulfate fractionation and several chromatographic steps, recombinant from Escherichia coli strain BL21(DE3) Methanopyrus kandleri

Source Tissue

Source Tissue Comment Organism Textmining
additional information optimal growth at 37°C Methanosarcina barkeri
-
additional information optimal growth at 65°C Methanothermobacter thermautotrophicus
-
additional information optimal growth at 83°C Archaeoglobus fulgidus
-
additional information optimal growth at 85°C Methanocaldococcus jannaschii
-
additional information optimal growth on H2 and CO2 at 98°C Methanopyrus kandleri
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Methanosarcina barkeri
additional information
-
-
Methanothermobacter thermautotrophicus
additional information
-
-
Archaeoglobus fulgidus
additional information
-
-
Methanopyrus kandleri

Storage Stability

Storage Stability Organism
-20°C, purified recombinant enzyme, N2-atmosphere, stable for at least 4 weeks Methanopyrus kandleri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methylenetetrahydromethanopterin + oxidized coenzyme F420 the enzyme is Si-face stereospecific with respect to C5 of F420 and Re-face stereospecific with respect to the methylene group of methylenetetrahydromethanopterin, reaction needs to be performed under strict anaerobic conditions Methanosarcina barkeri 5,10-methenyltetrahydromethanopterin + reduced coenzyme F420
-
r
5,10-methylenetetrahydromethanopterin + oxidized coenzyme F420 the enzyme is Si-face stereospecific with respect to C5 of F420 and Re-face stereospecific with respect to the methylene group of methylenetetrahydromethanopterin, reaction needs to be performed under strict anaerobic conditions Methanothermobacter thermautotrophicus 5,10-methenyltetrahydromethanopterin + reduced coenzyme F420
-
r
5,10-methylenetetrahydromethanopterin + oxidized coenzyme F420 the enzyme is Si-face stereospecific with respect to C5 of F420 and Re-face stereospecific with respect to the methylene group of methylenetetrahydromethanopterin, reaction needs to be performed under strict anaerobic conditions Methanocaldococcus jannaschii 5,10-methenyltetrahydromethanopterin + reduced coenzyme F420
-
r
5,10-methylenetetrahydromethanopterin + oxidized coenzyme F420 the enzyme is Si-face stereospecific with respect to C5 of F420 and Re-face stereospecific with respect to the methylene group of methylenetetrahydromethanopterin, reaction needs to be performed under strict anaerobic conditions Archaeoglobus fulgidus 5,10-methenyltetrahydromethanopterin + reduced coenzyme F420
-
r
5,10-methylenetetrahydromethanopterin + oxidized coenzyme F420 the enzyme is Si-face stereospecific with respect to C5 of F420 and Re-face stereospecific with respect to the methylene group of methylenetetrahydromethanopterin, reaction needs to be performed under strict anaerobic conditions Methanopyrus kandleri 5,10-methenyltetrahydromethanopterin + reduced coenzyme F420
-
r
additional information enzyme is involved in CO2 reduction to CH4 Methanosarcina barkeri ?
-
?
additional information enzyme is involved in CO2 reduction to CH4 Methanothermobacter thermautotrophicus ?
-
?
additional information enzyme is involved in CO2 reduction to CH4 Methanocaldococcus jannaschii ?
-
?
additional information enzyme is involved in CO2 reduction to CH4 Archaeoglobus fulgidus ?
-
?
additional information enzyme is involved in CO2 reduction to CH4 Methanopyrus kandleri ?
-
?

Subunits

Subunits Comment Organism
? x * 30305, sequence calculation Methanocaldococcus jannaschii
hexamer 6 * 36000, approximately, SDS-PAGE, 6 * 29644, sequence calculation Methanothermobacter thermautotrophicus
octamer 8 * 31000, approximately, SDS-PAGE Methanosarcina barkeri
octamer 8 * 36000, approximately, SDS-PAGE, 8 * 31383, sequence calculation Methanopyrus kandleri
tetramer 4 * 32000, approximately, SDS-PAGE, 4 * 29644, sequence calculation Archaeoglobus fulgidus

Synonyms

Synonyms Comment Organism
MTD
-
Methanosarcina barkeri
MTD
-
Methanothermobacter thermautotrophicus
MTD
-
Methanocaldococcus jannaschii
MTD
-
Archaeoglobus fulgidus
MTD
-
Methanopyrus kandleri
N5,N10-Methylenetetrahydromethanopterin dehydrogenase
-
Methanosarcina barkeri
N5,N10-Methylenetetrahydromethanopterin dehydrogenase
-
Methanothermobacter thermautotrophicus
N5,N10-Methylenetetrahydromethanopterin dehydrogenase
-
Methanocaldococcus jannaschii
N5,N10-Methylenetetrahydromethanopterin dehydrogenase
-
Archaeoglobus fulgidus
N5,N10-Methylenetetrahydromethanopterin dehydrogenase
-
Methanopyrus kandleri

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
-
Methanothermobacter thermautotrophicus
60
-
above Methanosarcina barkeri
70
-
-
Archaeoglobus fulgidus
75
-
-
Methanopyrus kandleri

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
90
-
30 min, stable up to over 90°C Methanopyrus kandleri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
assay at, substrate methylenetetrahydromethanopterin, forward reaction Methanopyrus kandleri
8
-
assay at, substrate methenyltetrahydromethanopterin, reverse reaction Methanopyrus kandleri

Cofactor

Cofactor Comment Organism Structure
coenzyme F420 dependent on Methanosarcina barkeri
coenzyme F420 dependent on Methanothermobacter thermautotrophicus
coenzyme F420 dependent on Methanocaldococcus jannaschii
coenzyme F420 dependent on Archaeoglobus fulgidus
coenzyme F420 dependent on Methanopyrus kandleri
additional information enzyme contains no prosthetic group Methanopyrus kandleri

pI Value

Organism Comment pI Value Maximum pI Value
Methanopyrus kandleri deduced from amino acid sequence
-
4.2
Methanothermobacter thermautotrophicus deduced from amino acid sequnce
-
4.9
Archaeoglobus fulgidus deduced from amino acid sequnce
-
4.9
Methanocaldococcus jannaschii deduced from amino acid sequnce
-
6