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Literature summary for 3.5.4.27 extracted from
- Tetrahydromethanopterin-specific enzymes from Methanopyrus kandleri (2001), Methods Enzymol., 331, 317-353.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene mch, DNA and amino acid sequence determination, overexpression in Escherichia coli strain BL21(DE3) | Methanopyrus kandleri |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant enzyme, X-ray diffraction structure determination and analysis at 2.0 A resolution | Methanopyrus kandleri |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.03 | - |
formyl-tetrahydromethanopterin | - |
Methylorubrum extorquens |  |
| 0.04 | - |
formyl-tetrahydromethanopterin | pH 8.0, 65°C | Methanopyrus kandleri | |
| 0.22 | - |
formyl-tetrahydromethanopterin | - |
Methanothermus fervidus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | lyotrophic salts at high concentrations above 1 M are required for enzyme activity, but not for stability, 200fold activation | Methanopyrus kandleri | |
| additional information | lyotrophic salts at high concentrations above 1 M are required for enzyme activity, but not for stability, 2fold activation | Methanothermus fervidus | |
| additional information | lyotrophic salts at high concentrations above 1 M are required for enzyme activity, but not for stability, 4fold activation | Methanosarcina barkeri | |
| additional information | lyotrophic salts at high concentrations above 1 M are required for enzyme activity, but not for stability, 65fold activation | Methanothermobacter thermautotrophicus | |
| additional information | lyotrophic salts at high concentrations above 1 M are required for enzyme activity, but not for stability, 7fold activation | Methylorubrum extorquens |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 33000 | - |
2 * 33282, sequence calculation, 2 * 33000, SDS-PAGE | Methylorubrum extorquens |
| 33282 | - |
2 * 33282, sequence calculation, 2 * 33000, SDS-PAGE | Methylorubrum extorquens |
| 33972 | - |
3 * 33972, sequence calculation, 3 * 41500, SDS-PAGE | Methanopyrus kandleri |
| 34246 | - |
2 * 34246, sequence calculation, 2 * 41000, SDS-PAGE | Methanothermobacter thermautotrophicus |
| 34851 | - |
2 * 34851, sequence calculation, 2 * 39000, SDS-PAGE | Methanothermus fervidus |
| 34889 | - |
2 * 34889, sequence calculation, 2 * 41000, SDS-PAGE | Methanosarcina barkeri |
| 34899 | - |
x * 34899, sequence calculation | Methanocaldococcus jannaschii |
| 39000 | - |
2 * 34851, sequence calculation, 2 * 39000, SDS-PAGE | Methanothermus fervidus |
| 41000 | - |
2 * 34246, sequence calculation, 2 * 41000, SDS-PAGE | Methanothermobacter thermautotrophicus |
| 41000 | - |
2 * 34889, sequence calculation, 2 * 41000, SDS-PAGE | Methanosarcina barkeri |
| 41500 | - |
3 * 33972, sequence calculation, 3 * 41500, SDS-PAGE | Methanopyrus kandleri |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| formyl-tetrahydromethanopterin | Methanosarcina barkeri | - |
methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | Methanothermobacter thermautotrophicus | - |
methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | Methanocaldococcus jannaschii | - |
methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | Methanothermus fervidus | - |
methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | Methanopyrus kandleri | - |
methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | Methylorubrum extorquens | - |
methenyl-tetrahydromethanopterin + H2O | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Methanocaldococcus jannaschii | - |
- |
- |
| Methanopyrus kandleri | - |
gene mch | - |
| Methanosarcina barkeri | - |
- |
- |
| Methanothermobacter thermautotrophicus | - |
- |
- |
| Methanothermus fervidus | - |
- |
- |
| Methylorubrum extorquens | - |
- |
- |
Oxidation Stability
| Oxidation Stability | Organism |
|---|---|
| the enzyme is slowly inactivated under oxic conditions | Methanopyrus kandleri |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation and ion exchange chromatography, native enzyme over 600fold from cell supernatant by ammonium sulfate fractionation and two steps of ion exchange chromatography, purification requires anaerobic conditions, the enzyme is completely soluble at 80% saturated ammonium sulfate | Methanopyrus kandleri |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| additional information | optimal growth temperature is 30°C | Methylorubrum extorquens | - |
| additional information | optimal growth temperature is 37°C | Methanosarcina barkeri | - |
| additional information | optimal growth temperature is 65°C | Methanothermobacter thermautotrophicus | - |
| additional information | optimal growth temperature is 83°C | Methanothermus fervidus | - |
| additional information | optimal growth temperature is 85°C | Methanocaldococcus jannaschii | - |
| additional information | optimal growth temperature is 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 | - |
- |
Methanothermus fervidus |
| additional information | - |
- |
Methylorubrum extorquens |
| 5000 | - |
above, purified native enzyme | Methanopyrus kandleri |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C, purified native enzyme, 1.32 M KCl, 50 mM Tris-HCl, pH 7.0, under N2 atmosphere, several months without significaant loss of activity | Methanopyrus kandleri |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| formyl-tetrahydromethanopterin | - |
Methanosarcina barkeri | methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | - |
Methanothermobacter thermautotrophicus | methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | - |
Methanocaldococcus jannaschii | methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | - |
Methanothermus fervidus | methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | - |
Methanopyrus kandleri | methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | - |
Methylorubrum extorquens | methenyl-tetrahydromethanopterin + H2O | - |
r | |
| formyl-tetrahydromethanopterin | substrate isolated and purified from Methanobacterium thermoautotrophicum, overview | Methanopyrus kandleri | methenyl-tetrahydromethanopterin + H2O | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 34899, sequence calculation | Methanocaldococcus jannaschii |
| dimer | 2 * 33282, sequence calculation, 2 * 33000, SDS-PAGE | Methylorubrum extorquens |
| dimer | 2 * 34246, sequence calculation, 2 * 41000, SDS-PAGE | Methanothermobacter thermautotrophicus |
| dimer | 2 * 34851, sequence calculation, 2 * 39000, SDS-PAGE | Methanothermus fervidus |
| dimer | 2 * 34889, sequence calculation, 2 * 41000, SDS-PAGE | Methanosarcina barkeri |
| trimer | 3 * 33972, sequence calculation, 3 * 41500, SDS-PAGE | Methanopyrus kandleri |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Mch | - |
Methanosarcina barkeri |
| Mch | - |
Methanothermobacter thermautotrophicus |
| Mch | - |
Methanocaldococcus jannaschii |
| Mch | - |
Methanothermus fervidus |
| Mch | - |
Methanopyrus kandleri |
| Mch | - |
Methylorubrum extorquens |
| N5,N10-methenyltetrahydromethanopterin cyclohydrolase | - |
Methanosarcina barkeri |
| N5,N10-methenyltetrahydromethanopterin cyclohydrolase | - |
Methanothermobacter thermautotrophicus |
| N5,N10-methenyltetrahydromethanopterin cyclohydrolase | - |
Methanocaldococcus jannaschii |
| N5,N10-methenyltetrahydromethanopterin cyclohydrolase | - |
Methanothermus fervidus |
| N5,N10-methenyltetrahydromethanopterin cyclohydrolase | - |
Methanopyrus kandleri |
| N5,N10-methenyltetrahydromethanopterin cyclohydrolase | - |
Methylorubrum extorquens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
optimal growth temperature of the organism is 30°C | Methylorubrum extorquens |
| additional information | - |
optimal growth temperature of the organism is 37°C | Methanosarcina barkeri |
| additional information | - |
optimal growth temperature of the organism is 65°C | Methanothermobacter thermautotrophicus |
| additional information | - |
optimal growth temperature of the organism is 83°C | Methanothermus fervidus |
| additional information | - |
optimal growth temperature of the organism is 85°C | Methanocaldococcus jannaschii |
| additional information | - |
optimal growth temperature of the organism is 98°C | Methanopyrus kandleri |
| 40 | - |
- |
Methylorubrum extorquens |
| 50 | - |
- |
Methanosarcina barkeri |
| 85 | - |
- |
Methanothermus fervidus |
| 95 | - |
- |
Methanopyrus kandleri |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
stable up to | Methanosarcina barkeri |
| 60 | - |
stable up to | Methylorubrum extorquens |
| 65 | - |
stable up to | Methanothermobacter thermautotrophicus |
| 90 | - |
stable up to | Methanothermus fervidus |
| 90 | - |
stable at 90°C and above | Methanopyrus kandleri |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Methanopyrus kandleri |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | the enzyme contains no prothetic group | Methanopyrus kandleri |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Methanopyrus kandleri | amino acid sequence calculation | - |
3.8 |
| Methanothermobacter thermautotrophicus | amino acid sequence calculation | - |
4.2 |
| Methanosarcina barkeri | amino acid sequence calculation | - |
4.3 |
| Methanothermus fervidus | amino acid sequence calculation | - |
4.3 |
| Methylorubrum extorquens | amino acid sequence calculation | - |
4.8 |
| Methanocaldococcus jannaschii | amino acid sequence calculation | - |
5.5 |
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