BRENDA - Enzyme Database
show all sequences of 1.5.99.15

Discovery and characterization of the first archaeal dihydromethanopterin reductase, an iron-sulfur flavoprotein from Methanosarcina mazei

Wang, S.; Tiongson, J.; Rasche, M.E.; J. Bacteriol. 196, 203-209 (2014)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
gene dmrX, recombinant expression of His6-tagged enzyme in Escherichia coli strain BE100 [BL21(DE3) RIL]
Methanosarcina mazei
gene dmrX, recombinant expression of the codon-optimized gene encoding His6-tagged enzyme in Escherichia col strain BE100 [BL21(DE3) RIL]
Methanocaldococcus jannaschii
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
the enzyme contains two [4Fe-4S] cluster sites
Methanosarcina mazei
Fe2+
the enzyme contains two [4Fe-4S] cluster sites
Methanocaldococcus jannaschii
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
7,8-dihydromethanopterin + reduced ferredoxin
Methanosarcina mazei
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
Methanocaldococcus jannaschii
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
Methanosarcina mazei DSM 3647
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
Methanocaldococcus jannaschii DSM 2661
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Methanocaldococcus jannaschii
Q57661
-
-
Methanocaldococcus jannaschii DSM 2661
Q57661
-
-
Methanosarcina mazei
Q8PVV3
-
-
Methanosarcina mazei DSM 3647
Q8PVV3
-
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant His6-tagged enzyme from Escherichia col strain BE100 [BL21(DE3) RIL] in an anaerobic chamber by heat treatement at 65°C for 5 min, the supernatant is a second time heat treated at 85°C for 15 min,
Methanocaldococcus jannaschii
recombinant His6-tagged enzyme from Escherichia coli strain BE100 [BL21(DE3) RIL] in an anaerobic chamber by nickel affinity chromatography, ultrafiltration, and gel filtration
Methanosarcina mazei
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanosarcina mazei
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanocaldococcus jannaschii
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanosarcina mazei DSM 3647
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanocaldococcus jannaschii DSM 2661
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MJ0208. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MJ0208 can transfer electrons to dihydromethanopterin
742781
Methanocaldococcus jannaschii
?
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MM1854. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MM1854 can transfer electrons to dihydromethanopterin
742781
Methanosarcina mazei
?
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MM1854. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MM1854 can transfer electrons to dihydromethanopterin
742781
Methanosarcina mazei DSM 3647
?
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MJ0208. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MJ0208 can transfer electrons to dihydromethanopterin
742781
Methanocaldococcus jannaschii DSM 2661
?
-
-
-
?
Subunits
Subunits
Commentary
Organism
homodimer
2 * 29000, recombinant His6-tagged enzyme, SDS-PAGE
Methanosarcina mazei
Synonyms
Synonyms
Commentary
Organism
AfpA
-
Methanocaldococcus jannaschii
archaeal-flavoprotein-like flavoprotein
-
Methanocaldococcus jannaschii
DmrX
-
Methanosarcina mazei
DmrX
-
Methanocaldococcus jannaschii
methylene H4MPT dehydrogenase B
-
Methanosarcina mazei
MJ0208
-
Methanocaldococcus jannaschii
MM1854
-
Methanosarcina mazei
MtdB
-
Methanosarcina mazei
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at room temperature
Methanosarcina mazei
22
-
assay at room temperature
Methanocaldococcus jannaschii
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
100
-
purified recombinant His-tagged enzyme, pH 7.0, 15 min, inactivation
Methanosarcina mazei
100
-
purified recombinant His-tagged enzyme, pH 7.0, 15 min, inactivation
Methanocaldococcus jannaschii
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
7.5
-
Methanosarcina mazei
7
7.5
-
Methanocaldococcus jannaschii
Cofactor
Cofactor
Commentary
Organism
Structure
Ferredoxin
ferredoxin may serve as an electron donor
Methanosarcina mazei
Ferredoxin
ferredoxin may serve as an electron donor
Methanocaldococcus jannaschii
FMN
the enzyme contains one flavin mononucleotide (FMN)-binding site
Methanosarcina mazei
FMN
the enzyme contains one flavin mononucleotide (FMN)-binding site
Methanocaldococcus jannaschii
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MJ0208. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MJ0208 can transfer electrons to dihydromethanopterin
Methanocaldococcus jannaschii
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MM1854. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MM1854 can transfer electrons to dihydromethanopterin
Methanosarcina mazei
[4Fe-4S]-center
the enzyme contains two iron-sulfur cluster sites
Methanosarcina mazei
[4Fe-4S]-center
the enzyme contains two iron-sulfur cluster sites
Methanocaldococcus jannaschii
Cloned(Commentary) (protein specific)
Commentary
Organism
gene dmrX, recombinant expression of His6-tagged enzyme in Escherichia coli strain BE100 [BL21(DE3) RIL]
Methanosarcina mazei
gene dmrX, recombinant expression of the codon-optimized gene encoding His6-tagged enzyme in Escherichia col strain BE100 [BL21(DE3) RIL]
Methanocaldococcus jannaschii
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
Ferredoxin
ferredoxin may serve as an electron donor
Methanosarcina mazei
Ferredoxin
ferredoxin may serve as an electron donor
Methanocaldococcus jannaschii
FMN
the enzyme contains one flavin mononucleotide (FMN)-binding site
Methanosarcina mazei
FMN
the enzyme contains one flavin mononucleotide (FMN)-binding site
Methanocaldococcus jannaschii
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MJ0208. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MJ0208 can transfer electrons to dihydromethanopterin
Methanocaldococcus jannaschii
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MM1854. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MM1854 can transfer electrons to dihydromethanopterin
Methanosarcina mazei
[4Fe-4S]-center
the enzyme contains two iron-sulfur cluster sites
Methanosarcina mazei
[4Fe-4S]-center
the enzyme contains two iron-sulfur cluster sites
Methanocaldococcus jannaschii
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
the enzyme contains two [4Fe-4S] cluster sites
Methanosarcina mazei
Fe2+
the enzyme contains two [4Fe-4S] cluster sites
Methanocaldococcus jannaschii
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
7,8-dihydromethanopterin + reduced ferredoxin
Methanosarcina mazei
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
Methanocaldococcus jannaschii
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
Methanosarcina mazei DSM 3647
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
Methanocaldococcus jannaschii DSM 2661
-
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His6-tagged enzyme from Escherichia col strain BE100 [BL21(DE3) RIL] in an anaerobic chamber by heat treatement at 65°C for 5 min, the supernatant is a second time heat treated at 85°C for 15 min,
Methanocaldococcus jannaschii
recombinant His6-tagged enzyme from Escherichia coli strain BE100 [BL21(DE3) RIL] in an anaerobic chamber by nickel affinity chromatography, ultrafiltration, and gel filtration
Methanosarcina mazei
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanosarcina mazei
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanocaldococcus jannaschii
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanosarcina mazei DSM 3647
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
7,8-dihydromethanopterin + reduced ferredoxin
-
742781
Methanocaldococcus jannaschii DSM 2661
5,6,7,8-tetrahydromethanopterin + ferredoxin
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MJ0208. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MJ0208 can transfer electrons to dihydromethanopterin
742781
Methanocaldococcus jannaschii
?
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MM1854. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MM1854 can transfer electrons to dihydromethanopterin
742781
Methanosarcina mazei
?
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MM1854. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MM1854 can transfer electrons to dihydromethanopterin
742781
Methanosarcina mazei DSM 3647
?
-
-
-
?
additional information
NAD(P)H is incapable of directly reducing the flavin cofactor, but dithionite eliminates the FMN peaks, indicating successful electron transfer to MJ0208. An electron transfer system of NADPH, spinach NADPH-ferredoxin oxidoreductase, and ferredoxin can also reduce the FMN peaks. Dithiothreitol-reduced MJ0208 can transfer electrons to dihydromethanopterin
742781
Methanocaldococcus jannaschii DSM 2661
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 29000, recombinant His6-tagged enzyme, SDS-PAGE
Methanosarcina mazei
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at room temperature
Methanosarcina mazei
22
-
assay at room temperature
Methanocaldococcus jannaschii
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
100
-
purified recombinant His-tagged enzyme, pH 7.0, 15 min, inactivation
Methanosarcina mazei
100
-
purified recombinant His-tagged enzyme, pH 7.0, 15 min, inactivation
Methanocaldococcus jannaschii
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
7.5
-
Methanosarcina mazei
7
7.5
-
Methanocaldococcus jannaschii
General Information
General Information
Commentary
Organism
metabolism
the microbial production of methane by methanogenic archaea is dependent on the synthesis of the pterin-containing cofactor tetrahydromethanopterin (H4MPT). The enzyme catalyzing the last step of H4MPT biosynthesis is dihydromethanopterin reductase
Methanosarcina mazei
metabolism
the microbial production of methane by methanogenic archaea is dependent on the synthesis of the pterin-containing cofactor tetrahydromethanopterin (H4MPT). The enzyme catalyzing the last step of H4MPT biosynthesis is dihydromethanopterin reductase
Methanocaldococcus jannaschii
physiological function
MJ0208 functions as an archaeal dihydromethanopterin reductase (DmrX) and ferredoxin may serve as an electron donor
Methanocaldococcus jannaschii
physiological function
MM1854 functions as an archaeal dihydromethanopterin reductase (DmrX) and ferredoxin may serve as an electron donor
Methanosarcina mazei
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the microbial production of methane by methanogenic archaea is dependent on the synthesis of the pterin-containing cofactor tetrahydromethanopterin (H4MPT). The enzyme catalyzing the last step of H4MPT biosynthesis is dihydromethanopterin reductase
Methanosarcina mazei
metabolism
the microbial production of methane by methanogenic archaea is dependent on the synthesis of the pterin-containing cofactor tetrahydromethanopterin (H4MPT). The enzyme catalyzing the last step of H4MPT biosynthesis is dihydromethanopterin reductase
Methanocaldococcus jannaschii
physiological function
MJ0208 functions as an archaeal dihydromethanopterin reductase (DmrX) and ferredoxin may serve as an electron donor
Methanocaldococcus jannaschii
physiological function
MM1854 functions as an archaeal dihydromethanopterin reductase (DmrX) and ferredoxin may serve as an electron donor
Methanosarcina mazei
Other publictions for EC 1.5.99.15
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
727782
Wang
Discovery and characterization ...
Methanocaldococcus jannaschii, Methanosarcina mazei, Methanosarcina mazei DSM 3647, Methanocaldococcus jannaschii DSM 2661
J. Bacteriol.
196
203-209
2014
-
-
2
-
-
1
-
-
-
2
1
4
-
17
-
-
2
-
-
-
-
1
8
1
4
-
-
-
-
-
-
-
2
-
-
-
-
-
2
2
-
-
1
-
-
-
-
-
2
1
4
-
-
-
2
-
-
-
1
8
1
-
-
-
-
-
-
-
-
-
2
2
-
-
-
742781
Wang
Discovery and characterizatio ...
Methanocaldococcus jannaschii, Methanosarcina mazei, Methanosarcina mazei DSM 3647, Methanocaldococcus jannaschii DSM 2661
J. Bacteriol.
196
203-209
2014
-
-
2
-
-
-
-
-
-
2
-
4
-
17
-
-
2
-
-
-
-
-
8
1
9
2
-
2
-
2
-
-
8
-
-
-
-
-
2
8
-
-
-
-
-
-
-
-
2
-
4
-
-
-
2
-
-
-
-
8
1
2
-
2
-
2
-
-
-
-
4
4
-
-
-
742870
McNamara
Structure of dihydromethanopt ...
Paraburkholderia xenovorans
J. Biol. Chem.
289
8852-8864
2014
-
-
2
2
-
-
-
-
-
-
-
2
-
1
-
-
2
1
-
-
-
-
3
2
4
-
-
-
-
-
-
-
3
-
-
-
-
-
2
3
2
-
-
-
-
-
-
-
-
-
2
-
-
-
2
-
-
-
-
3
2
-
-
-
-
-
-
-
-
-
3
3
-
-
-