Information on EC 1.5.99.15 - dihydromethanopterin reductase (acceptor)

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The expected taxonomic range for this enzyme is: Archaea, Bacteria

EC NUMBER
COMMENTARY hide
1.5.99.15
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RECOMMENDED NAME
GeneOntology No.
dihydromethanopterin reductase (acceptor)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
5,6,7,8-tetrahydromethanopterin + oxidized acceptor = 7,8-dihydromethanopterin + reduced acceptor
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
tetrahydromethanopterin biosynthesis
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methanogenesis from CO2
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Folate biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
5,6,7,8-tetrahydromethanopterin:acceptor 5,6-oxidoreductase
This archaeal enzyme catalyses the last step in the biosynthesis of tetrahydromethanopterin, a coenzyme used in methanogenesis. The enzyme, characterized from the archaea Methanosarcina mazei and Methanocaldococcus jannaschii, is an iron-sulfur flavoprotein. cf. EC 1.5.1.47, dihydromethanopterin reductase [NAD(P)+].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
sequence comparisons suggested that the catalytic mechanism is conserved among the bacterial homologues of DmrB and partially conserved in archaeal homologues, where an alternate electron donor is likely used
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
5,6,7,8-tetrahydromethanopterin + oxidized acceptor
7,8-dihydromethanopterin + reduced acceptor
show the reaction diagram
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?
7,8-dihydromethanopterin + FMNH2
5,6,7,8-tetrahydromethanopterin + FMN
show the reaction diagram
7,8-dihydromethanopterin + reduced acceptor
5,6,7,8-tetrahydromethanopterin + oxidized acceptor
show the reaction diagram
7,8-dihydromethanopterin + reduced dithiothreitol
5,6,7,8-tetrahydromethanopterin + oxidized dithiothreitol
show the reaction diagram
7,8-dihydromethanopterin + reduced ferredoxin
5,6,7,8-tetrahydromethanopterin + ferredoxin
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
5,6,7,8-tetrahydromethanopterin + oxidized acceptor
7,8-dihydromethanopterin + reduced acceptor
show the reaction diagram
Q13QT8
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?
7,8-dihydromethanopterin + FMNH2
5,6,7,8-tetrahydromethanopterin + FMN
show the reaction diagram
Q13QT8
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?
7,8-dihydromethanopterin + reduced acceptor
5,6,7,8-tetrahydromethanopterin + oxidized acceptor
show the reaction diagram
7,8-dihydromethanopterin + reduced ferredoxin
5,6,7,8-tetrahydromethanopterin + ferredoxin
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ferredoxin
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flavin
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iron-sulfur flavoprotein
[4Fe-4S]-center
additional information
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron-sulfur cluster
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
29000
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2 * 29000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homotetraicosamer
24 * 22100, His6-tagged enzyme, SDS-PAGE
homotrimer
enzyme crystal structure analysis
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 0.8-1.2 M ammonium sulfate as precipitant; purified recombinant N-terminally His6 -tagged enzyme by hanging drop vapor diffusion method, mixing of 0.003 ml of 13 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 5% glycerol, and 4 mM DTT, with reservori solution containing FMN, method optimization, 7-14 days at room temperature, X-ray diffraction structure determination and analysis at 1.9 A resolution
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
stable when frozen in liquid nitrogen
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
labile when exposed to cold storage at 4C and 20C or liquid nitrogen temperatures
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HisTrap column chromatography and S200 gel filtration; recombinant N-terminally His6 -tagged enzyme from Escherichia coli strain BL21DE3 RIL cell-free extract by nickel affinity chromatography and gel filtration
recombinant His6-tagged enzyme from Escherichia col strain BE100 [BL21(DE3) RIL] in an anaerobic chamber by heat treatement at 65C for 5 min, the supernatant is a second time heat treated at 85C for 15 min,
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recombinant His6-tagged enzyme from Escherichia coli strain BE100 [BL21(DE3) RIL] in an anaerobic chamber by nickel affinity chromatography, ultrafiltration, and gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21 (DE3)-RIL cells; gene dmrB, recombinant expression of N-terminally His6 -tagged enzyme in Escherichia coli strain BL21DE3 RIL
expression in Escherichia coli
gene dmrX, recombinant expression of His6-tagged enzyme in Escherichia coli strain BE100 [BL21(DE3) RIL]
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gene dmrX, recombinant expression of the codon-optimized gene encoding His6-tagged enzyme in Escherichia col strain BE100 [BL21(DE3) RIL]
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