Information on EC 3.1.3.77 - acireductone synthase

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

EC NUMBER
COMMENTARY hide
3.1.3.77
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RECOMMENDED NAME
GeneOntology No.
acireductone synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-hydroxy-5-(methylthio)-3-oxopent-1-enyl phosphate + H2O = 1,2-dihydroxy-5-(methylthio)pent-1-en-3-one + phosphate
show the reaction diagram
(1b)
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5-(methylthio)-2,3-dioxopentyl phosphate + H2O = 1,2-dihydroxy-5-(methylthio)pent-1-en-3-one + phosphate
show the reaction diagram
overall reaction
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5-(methylthio)-2,3-dioxopentyl phosphate = 2-hydroxy-5-(methylthio)-3-oxopent-1-enyl phosphate
show the reaction diagram
(1a)
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
methionine metabolism
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Cysteine and methionine metabolism
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
5-(methylthio)-2,3-dioxopentyl-phosphate phosphohydrolase (isomerizing)
This bifunctional enzyme first enolizes the substrate to form the intermediate 2-hydroxy-5-(methylthio)-3-oxopent-1-enyl phosphate, which is then dephosphorylated to form the acireductone 1,2-dihydroxy-5-(methylthio)pent-1-en-3-one [2]. The acireductone represents a branch point in the methione-salvage pathway as it is used in the formation of formate, CO and 3-(methylthio)propanoate by EC 1.13.11.53 [acireductone dioxygenase (Ni2+-requiring)] and of formate and 4-methylthio-2-oxobutanoate either by a spontaneous reaction under aerobic conditions or by EC 1.13.11.54 {acireductone dioxygenase [iron(II)-requiring]} [1,2].
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
methionine salvage pathway
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(1-oxo-heptyl)-phosphonic acid + H2O
?
show the reaction diagram
substrate analog, in complex with MASA it sits in the cleft formed by the conserved residues in the active site
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?
2,3-diketo-1-phosphohexane + H2O
2-ketopentanoate + formate + glyoxylate + butyrate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.077
2,3-Diketo-1-phosphohexane
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0.0071
2-hydroxy-3-keto-1-phospho-1-hexene
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.001
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clone MM294, transformant DIOX1, DIOX3, DIOX4 and non-hybridizing isolate taken as a negative control
0.00816
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UNF932 strain
0.179
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clone MM294, transformant DIOX2
21
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purified enzyme by phenyl-Superose column
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25500
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gel filtration
25620
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deduced from amino acid sequence
27000
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calculated from sequence
28300
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SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by hanging drop vapour-diffusion method, to 1.7 A resolution, crystals belong to space group P212121, with unit-cell parameters a = 54.02 A, b = 57.55 A, c = 87.32 A
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crystal structure to 1.7 A resolution of MASA and its complex with a substrate analog, by hanging-drop vapor-diffusion technique
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, 20 mM HEPES buffer, pH 7.5
-80°C, 50% glycerol
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
100fold purified to homogeneity in 10% yield, SDS-PAGE, amino acid sequence analysis
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by Ni2+-chelating affinity column, gel filtration and anion-exchange chromatography, more than 90% pure
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gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
for genetic complementation
into the bacterial expression vector pET28a with a His tag on the C-terminus. Recombinant plasmid overexpressed in Escherichia coli strain BL21 (DE3)
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Show AA Sequence (2501 entries)
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