Information on EC 4.1.1.3 - Oxaloacetate decarboxylase

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

EC NUMBER
COMMENTARY
4.1.1.3
-
RECOMMENDED NAME
GeneOntology No.
Oxaloacetate decarboxylase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
oxaloacetate = pyruvate + CO2
show the reaction diagram
-
-
-
-
oxaloacetate = pyruvate + CO2
show the reaction diagram
Keq: 0.00213 1/mM, decarboxylation favored; pyruvate + CO2 = oxaloacetae
-
oxaloacetate = pyruvate + CO2
show the reaction diagram
not
-
oxaloacetate = pyruvate + CO2
show the reaction diagram
pyruvate + CO2 = oxaloacetae
-
oxaloacetate = pyruvate + CO2
show the reaction diagram
pyruvate + CO2 = oxaloacetae
-
oxaloacetate = pyruvate + CO2
show the reaction diagram
catalytic mechanism, overview. Carboxybiotin transits to the membrane-bound beta-subunit where it is decarboxylated to biotin and CO2 in a reaction that consumes a periplasmic proton and is coupled to Na+ translocation from the cytoplasm to the periplasm. The reaction is initiated by the enzyme-catalyzed decarboxylation of oxaloacetate in the carboxyltransferase domain of the alpha-subunit, yielding pyruvate and carboxybiotin. Subsequently, the C-terminal biotin carboxyl carrier protein domain on the alpha-subunit translocates to the beta-subunit where its decarboxylation is coupled to Na+ translocation. The OADC pump is reversible: at high concentrations of extracellular Na+, the pump will couple the downhill movement of Na+ into the cytosol with the carboxylation of pyruvate, to form oxaloacetate
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
carboxylation
-
-
-
-
decarboxylation
-
-
-
-
decarboxylation
-, Q9HUU1
-
PATHWAY
KEGG Link
MetaCyc Link
methylgallate degradation
-
protocatechuate degradation I (meta-cleavage pathway)
-
syringate degradation
-
Pyruvate metabolism
-
Metabolic pathways
-
SYSTEMATIC NAME
IUBMB Comments
oxaloacetate carboxy-lyase (pyruvate-forming)
The enzyme from Klebsiella aerogenes is a biotinyl protein and also decarboxylates glutaconyl-CoA and methylmalonyl-CoA. The process is accompanied by the extrusion of two sodium ions from cells. Some animal enzymes require Mn2+.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Cg1458 protein
-
-
OAD-1
Q6A1G0
-
OAD-1
Vibrio cholerae O395-N1
Q6A1G0
-
-
OAD-2
Q6A1F6
-
OAD-2
Vibrio cholerae O395-N1
Q6A1F6
-
-
Oxalacetate decarboxylase
-
-
-
-
Oxalacetic acid decarboxylase
-
-
-
-
Oxalacetic beta-decarboxylase
-
-
-
-
Oxalacetic carboxylase
-
-
-
-
Oxalate beta-decarboxylase
-
-
-
-
oxaloacetate carboxylase Na+ pump
-
membrane-bound multiprotein complex that couples oxaloacetate decarboxylation to sodium ion transport across the membrane
Oxaloacetate carboxylyase
-
-
-
-
oxaloacetate decarboxylase
-
-
oxaloacetate decarboxylase Na+ pump
Q6A1F6, Q6A1G0
-
oxaloacetate decarboxylase Na+ pump
Vibrio cholerae O395-N1
Q6A1F6, Q6A1G0
-
-
oxaloacetate decarboxylase Na+ pump OAD-1
-
-
oxaloacetate decarboxylase Na+ pump OAD-2
-
-
oxaloacetate decarboxylase OAD-1
Q6A1G0
-
oxaloacetate decarboxylase OAD-1
Vibrio cholerae O395-N1
Q6A1G0
-
-
oxaloacetate decarboxylase OAD-2
Q6A1F6
-
oxaloacetate decarboxylase OAD-2
Vibrio cholerae O395-N1
Q6A1F6
-
-
PA4872
-
member of the PEP mutase/isocitrate lyase superfamily
CAS REGISTRY NUMBER
COMMENTARY
9024-98-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
guinea pig
-
-
Manually annotated by BRENDA team
strain ATCC 13032
-
-
Manually annotated by BRENDA team
genes oadA, oadB, oadC, and oadD
-
-
Manually annotated by BRENDA team
strain ATCC 29212
-
-
Manually annotated by BRENDA team
genes oadA, oadB, oadC, and oadD
-
-
Manually annotated by BRENDA team
Gluconacetobacter xylinus
-
-
-
Manually annotated by BRENDA team
sunflower
-
-
Manually annotated by BRENDA team
subspecies Lactococcus lactis lactis, strain IL1403
-
-
Manually annotated by BRENDA team
Lactococcus lactis CRL264
CRL264
-
-
Manually annotated by BRENDA team
formerly Micrococcus lysodeiktikus
-
-
Manually annotated by BRENDA team
pig
-
-
Manually annotated by BRENDA team
oxaloacetate decarboxylase OAD-1, alpha subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
oxaloacetate decarboxylase OAD-1, beta subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
oxaloacetate decarboxylase OAD-1, gamma subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
oxaloacetate decarboxylase OAD-2, alpha subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
oxaloacetate decarboxylase OAD-2, beta subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
oxaloacetate decarboxylase OAD-2, gamma subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
Vibrio cholerae O395-N1
oxaloacetate decarboxylase OAD-1, alpha subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
Vibrio cholerae O395-N1
oxaloacetate decarboxylase OAD-1, beta subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
Vibrio cholerae O395-N1
oxaloacetate decarboxylase OAD-1, gamma subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
Vibrio cholerae O395-N1
oxaloacetate decarboxylase OAD-2, alpha subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
Vibrio cholerae O395-N1
oxaloacetate decarboxylase OAD-2, beta subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
Vibrio cholerae O395-N1
oxaloacetate decarboxylase OAD-2, gamma subunit; O395-N1
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
the enzyme belongs to the class II decarboxylases of the biotin-dependent enzyme family. Class II enzymes facilitate sodium transport from the cytoplasm to the periplasm in some archaea and anaerobic bacteria
malfunction
-
inactivation of the odx gene does not improve L-lysine production
malfunction
Q82YW6
the citrate fermentation phenotype is not affected by citM deletion
metabolism
Q82YW6
CitM is not required for efficient citrate utilization
physiological function
-
presence of the membrane-bound Ef-B subunit is required for full alkalinization of the internal medium of Enterococcus faecalis cells during citrate fermentation
physiological function
-
the membrane-bound oxaloacetate decarboxylase complex of Klebsiella aerogenes catalyzes the biotin-dependent decarboxylation of oxaloacetate, while also serves as a primary Na+ pump. The enzyme complex plays an essential role in the citrate or tartrate fermentation pathways of certain archaea and bacteria, contributing to the generation of an electrochemical gradient of Na+ ions along with one mol of ATP per mol of citrate/tartrate. The resulting Na+ gradient is used to power the import of nutrients and the synthesis of ATP
physiological function
-
presence of the membrane-bound Ef-B subunit is required for full alkalinization of the internal medium of Enterococcus faecalis cells during citrate fermentation
-
metabolism
-
OAD of Vibrio cholerae catalyses a key step in citrate fermentation, converting the chemical energy of the decarboxylation reaction into an electrochemical gradient of Na+ ions across the membrane, which drives endergonic membrane reactions such as ATP synthesis, transport and motility
additional information
-
oxaloacetate decarboxylase complex structure, modeling, overview. The gamma-subunit is essential for the overall stability of the complex, and likely serves as an anchor to hold the alpha- and beta-subunits in place. The gamma-subunit significantly accelerates the rate of oxaloacetate decarboxylation in the alpha-subunit, which correlates with the coordination of a Zn2+ metal ion by several residues at the hydrophilic C-terminus. The 65 kDa hydrophilic alpha-subunit consists of an N-terminal carboxyltransferase domain connected to a C-terminal biotin carboxyl carrier protein domain. The 45 kDa beta-subunit is an integral membrane protein with nine transmembrane segments, which serves to couple the decarboxylation of carboxybiotin to the translocation of Na+ from the cytoplasm to the periplasm. The small 9 kDa gamma-subunit is an integral membrane protein with a single membrane-spanning helix at the N-terminus, followed by a hydrophilic C-terminal domain which interacts with the alpha-subunit. The gamma-subunit is essential for the overall stability of the complex, and likely serves as an anchor to hold the alpha- and beta-subunits in place
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-, Q9HUU1
-
-
-
r
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
ir
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
ir
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
Gluconacetobacter xylinus
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
r
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
Q82YW6
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
reaction mechanism
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
reaction mechanism
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
reaction mechanism
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
stereochemistry
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
reaction is catalyzed at low rate by the alpha subunit alone, much higher activity with the complete enzyme
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
two Na+ are transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
Lactococcus lactis CRL264
-
-
-
-
ir
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
Gluconacetobacter xylinus
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
part of citrate fermentation pathway, amount of Na+ transported depends on decarboxylation rate, but decoupling at very high decarboxylation rates, electrochemical gradient equals membrane driving force of 114 mV
-
-
-
oxaloacetate
?
show the reaction diagram
-
high activity in ethanol- or acetate-grown bacteria
-
-
-
oxaloacetate
?
show the reaction diagram
-
decarboxylation energy used for active Na+ transport
-
-
-
oxaloacetate
?
show the reaction diagram
-
decarboxylation energy used for active Na+ transport
-
-
-
oxaloacetate
?
show the reaction diagram
-
Na+ transport coupled to oxaloacetate decarboxylation
-
-
-
oxaloacetate
?
show the reaction diagram
-
involved in citrate fermentation
-
-
-
oxaloacetate
?
show the reaction diagram
-
involved in citrate fermentation
-
-
-
Pyruvate + CO2
Oxaloacetate
show the reaction diagram
-
-
-
-
Pyruvate + CO2
Oxaloacetate
show the reaction diagram
-
-
-
-
Pyruvate + CO2
Oxaloacetate
show the reaction diagram
-
-
-
-
Pyruvate + CO2
Oxaloacetate
show the reaction diagram
-
not
-
-
-
Pyruvate + CO2
Oxaloacetate
show the reaction diagram
-
reversal of reaction with whole enzyme by appropriate transmembrane Na+ gradient
-
-
3-methyloxaloacetate
3-methylpyruvate + CO2
show the reaction diagram
-, Q9HUU1
-
-
-
?
additional information
?
-
-
transmembrane transport of Na+
-
-
-
additional information
?
-
-
transmembrane transport of Na+
-
-
-
additional information
?
-
-
transmembrane transport of Na+
-
-
-
additional information
?
-
-
no substrate: phosphoenolpyruvate
-
-
-
additional information
?
-
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
after reconstitution into proteoliposomes, the enzyme acts as a Na+ pump
-
-
-
additional information
?
-
-
decarboxylation and sodium transport by the biotin-dependent oxaloacetate decarboxylase complex, overview
-
-
-
additional information
?
-
Vibrio cholerae O395-N1
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1
after reconstitution into proteoliposomes, the enzyme acts as a Na+ pump
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
one Na+ is transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
two Na+ are transported into the periplasm and one H+ is transported into the cytoplasm during the reaction
-
?
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
Gluconacetobacter xylinus
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
-
-
-
-
oxaloacetate
?
show the reaction diagram
-
part of citrate fermentation pathway, amount of Na+ transported depends on decarboxylation rate, but decoupling at very high decarboxylation rates, electrochemical gradient equals membrane driving force of 114 mV
-
-
-
oxaloacetate
?
show the reaction diagram
-
high activity in ethanol- or acetate-grown bacteria
-
-
-
oxaloacetate
?
show the reaction diagram
-
decarboxylation energy used for active Na+ transport
-
-
-
oxaloacetate
?
show the reaction diagram
-
decarboxylation energy used for active Na+ transport
-
-
-
oxaloacetate
?
show the reaction diagram
-
Na+ transport coupled to oxaloacetate decarboxylation
-
-
-
oxaloacetate
?
show the reaction diagram
-
involved in citrate fermentation
-
-
-
oxaloacetate
?
show the reaction diagram
-
involved in citrate fermentation
-
-
-
Oxaloacetate
Pyruvate + CO2
show the reaction diagram
-
-
-
-
?
additional information
?
-
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
after reconstitution into proteoliposomes, the enzyme acts as a Na+ pump
-
-
-
additional information
?
-
-
decarboxylation and sodium transport by the biotin-dependent oxaloacetate decarboxylase complex, overview
-
-
-
additional information
?
-
Vibrio cholerae O395-N1
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1
after reconstitution into proteoliposomes, the enzyme acts as a Na+ pump
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
biotin
-
about 1 mol bound per alpha-chain
biotin
-
role in enzyme function
biotin
-
binds to the C-terminal domain of the alpha-subunit
NAD+
Gluconacetobacter xylinus
-
reaction enhancement, allosteric effector
NAD+
-
one pyridine nucleotide reduced for each pyruvate formed
NADP+
Gluconacetobacter xylinus
-
reaction enhancement
NADP+
-
one pyridine nucleotide reduced for each pyruvate formed
biotin
-
dependent on, the enzyme utilizes a carboxyltransferase domain to catalyze the biotin-dependent decarboxylation of oxaloacetate
additional information
Q82YW6
no nicotinamide cofactors required
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Cd2+
-
activates
Co2+
-
activates
Co2+
-
activates
Li+
-
Km: 25 mM
Mg2+
-
required, decarboxylation Km: 0.17 mM, carboxylation Km: 1.85 mM
Mg2+
Gluconacetobacter xylinus
-
Km: 1.2 mM, can be substituted by Mn2+
Mg2+
-
only cytoplasmic enzyme
Mg2+
-
activates
Mg2+
-
inhibitory above 1 mM
Mg2+
-
or Mn2+, absolutely required, Km-value 0.08 mM
Mg2+
-
activates
Mg2+
-, Q9HUU1
cofactor
Mn2+
-
required
Mn2+
Gluconacetobacter xylinus
-
can be substituted by Mg2+, Km: 0.08 mM
Mn2+
-
only cytoplasmic enzyme
Mn2+
-
activates
Mn2+
-
Km: 1.2 mM; required, can be substituted by Mg2+, Co2+, Ni2+, Ca2+
Mn2+
-
required, can be substituted by Mg2+, Ca2+
Mn2+
-
inhibitory above 1 mM
Mn2+
-
or Mg2+, absolutely required, Km-value 0.02 mM, inhibitory above 0.46 mM
Mn2+
-
dependent on, maximal activity in presence of 10 mM Mn2+
Mn2+
Q82YW6
the enzyme is dependent on divalent metal ions with 100% activity at 20 mM Mn2+
Na+
-
Km: 1 mM
Na+
-
Km: 1.5 mM
Na+
-
Km: 0.78 mM
Na+
-
protects against proteolysis and alcohol inactivation
Na+
-
stoichiometry with decarboxylation, Na+ transport directly depends on biotin carboxylation/decarboxylation
Na+
-
two ions per enzyme required for maximum activity, cooperative binding
Na+
-
two ions per enzyme required for maximum activity, cooperative binding, 20-50 mM protects against tryptic digestion
Na+
-
protects against tryptic digestion
Na+
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
dependent on
Ni2+
-
activates
Ni2+
-
activates
Zn2+
-
gamma-subunit, involved in carboxytransfer, 0.96 mol/mol enzyme
Zn2+
-
0.61-0.99 mol/mol enzyme
Zn2+
-
binds to the gamma subunit, can not be removed with EDTA or 1,10-phenanthroline
Zn2+
-
binds to the gamma subunit
Zn2+
-
active site metal ion, 2 mol Zn2+ per mol enzyme
Zn2+
-
subunit Oad-gamma contains a Zn(II)-bound metal
Zn2+
-
the gamma subunit contains zinc
Zn2+
-
bound at the gamma-subunit, coordinated by several residues at the hydrophilic C-terminus
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-oxoglutarate
-
2 mM inhibitor in presence of 10 mM Mn2+, 11% inhibition
2-Oxomalonate
-
Ki: 0.06 mM
3,3-difluoroxaloacetate
-, Q9HUU1
-
5,5'-dithiobis(2-nitrobenzoate)
-
-
acetate
Gluconacetobacter xylinus
-
competitive for oxaloacetate, Ki: 12 mM
acetic acid
-
noncompetitive
Acetopyruvate
-, Q9HUU1
-
ADP
-
Ki: 1.5 mM
ADP
Q82YW6
78% residual activity at 0.25 mM
alpha-ketovalerate
-, Q9HUU1
-
ATP
-
carboxylation competitively inhibited, decarboxylation non-competitively, Ki: 2.2 mM, possibly involved in regulation
ATP
-
Ki: 1.5 mM
ATP
-
complete inhibition at 8-9 mM
ATP
Q82YW6
37% residual activity at 1 mM
Avidin
-
not
-
azide
-
2 mM, 20% inhibition
Ca2+
-
2 mM inhibitor in presence of 1 mM Mn2+, 30% inhibition
citrate
-
50% inhibition of mitochondrial enzyme at 4.5 mM
citrate
-
-
citrate
-
0.5 mM inhibitor or 2 mM inhibitor in presence of 10 mM Mn2+, 24% inhibition
citrate
Q82YW6
74% residual activity at 2 mM
coenzyme A
-
50% inhibition of mitochondrial enzyme at 0.05 mM
coenzyme A
-
Ki: 2.4 mM
Cu2+
-
2 mM, 45% inhibition
Cu2+
-
0.5 mM inhibitor in presence of 1 mM Mn2+, complete inhibition
Diethylstilbestrol
-
95% inhibition at 50 mM
EDTA
Gluconacetobacter xylinus
-
inhibition of residual activity without divalent metal ions
EDTA
-
2 mM, 100% inhibition
EDTA
-
10 mM, complete inhibition
EDTA
Q82YW6
complete inhibition at 2 mM
fumarate
-
2 mM inhibitor in presence of 10 mM Mn2+, 21% inhibition
glyoxylate
-
Ki: 0.04 mM
Hg2+
-
complete inactivation at 1 mM, buffer-dependend, interaction with alpha-subunit
hydrogencarbonate
-
-
KSCN
-
95% inhibition at 50 mM, completely reversible
L-Malate
Gluconacetobacter xylinus
-
competitive for oxaloacetate, Ki: 4.5 mM
malate
-
2 mM inhibitor in presence of 10 mM Mn2+, 54% inhibition
malate
Q82YW6
65% residual activity at 2 mM
malic acid
-
competitive
malonate
-
-
malonate
-
0.5 mM inhibitor in presence of 10 mM Mn2+, 84% inhibition
malonate
Q82YW6
26% residual activity at 2 mM
Mn2+
Gluconacetobacter xylinus
-
at concentrations higher than 0.4 mM
N-ethylmaleimide
-
-
N-ethylmaleimide
-
2 mM, 14% inhibition
NaCl
-
almost complete inhibition at 0.4 M
NaCl
-
OAD tertiary structure is sensitive to the presence of NaCl
NAD+
-
2 mM inhibitor in presence of 10 mM Mn2+, complete inhibition
NAD+
Q82YW6
40% residual activity at 0.25 mM
NADH
-
2 mM inhibitor in presence of 10 mM Mn2+, complete inhibition
NADH
Q82YW6
53% residual activity at 0.25 mM
Ni2+
-
2 mM inhibitor in presence of 1 mM Mn2+, 40% inhibition
oxalate
-
competitive for oxaloacetate, Ki: 0.0038 mM
oxalate
-
Ki: 0.0035
oxalate
-
Ki: 0.005 mM
oxalate
-
-
oxalate
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
half-maximal inhibition at 0.01 mM
oxalate
-
0.5 mM inhibitor in presence of 10 mM Mn2+, 80% inhibition
oxalate
Q82YW6
14% residual activity at 2 mM
oxalic acid
-
competitive
oxomalonate
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
half-maximal inhibition at 0.2 mM
oxomalonate
-
competitive inhibitor of OAD with respect to oxaloacetate
p-chloromercuribenzoate
-
-
p-chloromercuribenzoate
-
50% inhibition at 0.5 mM
p-chloromercuribenzoate
-
-
p-chloromercuribenzoate
-
2 mM, 62% inhibition
p-hydroxymercuribenzoate
-
70% inhibition at 5 mM
p-hydroxymercuribenzoate
Gluconacetobacter xylinus
-
complete inactivation after preincubation at 0.2 mM
p-hydroxymercuribenzoate
-
partly prevented by NADP+
phosphoenolpyruvate
-
50% inhibition of mitochondrial enzyme at 28 mM
phosphoenolpyruvate
-
competitive
Phosphonopyruvate
-, Q9HUU1
-
pyruvate
-
product inhibition competitive for oxaloacetate, Ki: 1.3 mM, noncompetitive inhibition for Na+, Ki: 2 mM
pyruvate
-
2 mM inhibitor in presence of 10 mM Mn2+, 8% inhibition
pyruvate
-, Q9HUU1
-
succinate
-
2 mM inhibitor in presence of 10 mM Mn2+, 23% inhibition
Zn2+
-
54% inhibition at 1.5 mM
Zn2+
-
0.5 mM inhibitor in presence of 1 mM Mn2+, complete inhibition
Mn2+
-
inhibitory above 0.46 mM
additional information
-
no effect is observed after addition of ADP (2.4 mM), GDP (3.6 mM), or succinate (5.6 mM) or after addition of phosphoenolpyruvate, L-fructose-6-phosphate, L-fructose-1,6-bisphosphate, L-malate, acetate, citrate, fumarate, L-aspartate, or acetyl coenzyme A (5 mM each)
-
additional information
Q82YW6
not inhibited by succinate
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
citrate
-
expression is induced by citrate
EDTA
-
activating below 5 mM
NaCl
-
highest activity with 1-1.4 mM
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.63
-
3-Methyloxaloacetate
-, Q9HUU1
wild type enzyme, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25C
0.0003
-
oxaloacetate
-
wild-type, Vmax = 4 micromol/min/mg; Y235A mutant, Vmax = 0.2 micromol/min/mg. In the presence of GDP; Y235F mutant, Vmax = 2 micromol/min/mg; Y235S mutant, Vmax = 0.1 micromol/min/mg. In the presence of GDP
0.0004
-
oxaloacetate
-
wild-type, Vmax = 3 micromol/min/mg. In the presence of GDP; Y235F mutant, Vmax = 3 micromol/min/mg. In the presence of GDP
0.03
-
oxaloacetate
-
pH 7.1
0.06
-
oxaloacetate
-
-
0.087
-
oxaloacetate
-, Q9HUU1
mutant enzyme Y212F, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25C
0.15
-
oxaloacetate
-
-
0.15
-
oxaloacetate
-
-
0.22
-
oxaloacetate
-
-
0.23
-
oxaloacetate
-
mitochondrial enzyme
0.52
-
oxaloacetate
-
-
0.55
-
oxaloacetate
-
-
0.62
-
oxaloacetate
Q82YW6
calculated value at pH 4.5, temperature not specified in the publication
1.02
-
oxaloacetate
-, Q9HUU1
mutant enzyme H235A, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25C
1.39
-
oxaloacetate
-
in 100 mM Tris-HCl (pH 7.3), 10 mM MgCl2, 2.5 mM EDTA, at 30C
1.7
-
oxaloacetate
-
-
2.1
-
oxaloacetate
-
-
2.1
-
oxaloacetate
-
pH 8.0, 25C
2.2
-
oxaloacetate
-, Q9HUU1
wild type enzyme, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25C
2.5
-
oxaloacetate
-, Q9HUU1
mutant enzyme H235Q, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25C
4.3
-
oxaloacetate
-
-
3.3
-
pyruvate
-
-
additional information
-
additional information
Gluconacetobacter xylinus
-
kinetic data; positive cooperativity for oxaloacetate
-
additional information
-
additional information
-
kinetic data
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
250
-
3-Methyloxaloacetate
-, Q9HUU1
in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25C
3.5
-
oxaloacetate
-
-
11.2
-
oxaloacetate
Q82YW6
calculated value at pH 4.5, temperature not specified in the publication
20.26
-
oxaloacetate
-
-
104
-
oxaloacetate
-
in 100 mM Tris-HCl (pH 7.3), 10 mM MgCl2, 2.5 mM EDTA, at 30C
7500
-
oxaloacetate
-, Q9HUU1
in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
22.3
-
oxaloacetate
Q82YW6
calculated value at pH 4.5, temperature not specified in the publication
57
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.06
-
2-Oxomalonate
-
-
0.45
-
3,3-difluoroxaloacetate
-, Q9HUU1
-
18.4
-
acetic acid
-
pH 8.0, 25C
1.09
-
Acetopyruvate
-, Q9HUU1
-
5.1
-
ADP
-
pH 8.0, 25C
6.7
-
alpha-ketovalerate
-, Q9HUU1
-
0.04
-
glyoxylate
-
-
2.2
-
hydrogencarbonate
-
pH 8.0, 25C
21.4
-
malic acid
-
pH 8.0, 25C
198
-
NaCl
-
wild type enzyme, pH 6.9
0.0035
-
oxalate
-
-
0.005
-
oxalate
-
-
0.043
-
oxalate
-, Q9HUU1
-
0.6
-
oxalic acid
-
pH 8.0, 25C
0.5
-
p-chloromercuribenzoate
-
-
7.8
-
phosphoenolpyruvate
-
pH 8.0, 25C
28
-
phosphoenolpyruvate
-
mitochondrial enzyme
3
-
Phosphonopyruvate
-, Q9HUU1
-
1.3
-
pyruvate
-
product inhibition competitive for oxaloacetate
2.1
-
pyruvate
-
pH 8.0, 25C
7.2
-
pyruvate
-, Q9HUU1
-
2.5
-
Mn2+
-
pH 8.0, 25C
additional information
-
additional information
-
Ki-values given for inhibition with NaCl for all mutant enzymes at 3-4 different pH
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.54
-
-
-
3.15
-
-
purified enzyme, pH 7.1
9
-
-
alpha-chain, independent of Na+
45
-
-
wild type enzyme
319
-
-
pH 8.0, 25C
360
-
Gluconacetobacter xylinus
-
-
additional information
-
-
-
additional information
-
Gluconacetobacter xylinus
-
-
additional information
-
-
-
additional information
-
-
activity of reconstituted enzyme and isolated subunits
additional information
-
-
strain-dependend
additional information
-
-
specific activity for all mutant enzymes
additional information
-
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
-
Q82YW6
-
5
-
-
cytoplasmic enzyme
5.5
-
-
Q192L and Y209A mutants
6
-
Gluconacetobacter xylinus
-
enzyme assay at
6.25
6.75
-
wild type enzyme
6.5
7
-
wild type enzyme
6.5
7.5
-
-
6.5
7.5
-
-
6.5
7.5
-
wild type enzyme
7
-
-
enzyme assay at
7
-
-
-
7
-
-
pH optimum at about pH 7 for all mutant enzymes except R389A
7.1
-
-
-
8.5
-
-
pH-optima above 8.5 found for N373L, R389A, R389L mutant enzymes
9.2
-
-
R389A mutant
9.5
-
-
mitochondrial enzyme
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.5
5
Q82YW6
-
3.5
5.5
-
pH 3.5: about 55% of maximal activity, pH 5.5: about 40% of maximal activity
6
8
-
-
6
8.5
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22
-
Gluconacetobacter xylinus
-
enzyme assay at
25
-
-
enzyme assay at
30
-
-
enzyme assay at
30
-
-
enzyme assay at
37
-
-
enzyme assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
58
-
-
no activity above
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
upon anaerobic growth of Vibrio cholerae on citrate, exclusivle the oad-2 gene is expressed
Manually annotated by BRENDA team
Vibrio cholerae O395-N1
-
upon anaerobic growth of Vibrio cholerae on citrate, exclusivle the oad-2 gene is expressed
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
83% of activity
Manually annotated by BRENDA team
Gluconacetobacter xylinus
-
-
Manually annotated by BRENDA team
-
differs from mitochondrial enzyme
Manually annotated by BRENDA team
-
isolated alpha-subunits or reconstituted alpha-beta-complexes expressed in Escherichia coli
Manually annotated by BRENDA team
-
subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex, Ef-AHD, which is also associated with the membrane
Manually annotated by BRENDA team
-
subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex, Ef-AHD, which is also associated with the membrane
-
Manually annotated by BRENDA team
-
alpha-subunit is peripheral membrane protein, beta- and gamma-subunits are integral membrane proteins, spatial model
Manually annotated by BRENDA team
-
reconstituted gamma-alpha-subunits or whole enzyme expressed in Escherichia coli
Manually annotated by BRENDA team
-
beta and gamma subunits are integral membrane proteins, alpha subunit is attached to the gamma subunit
Manually annotated by BRENDA team
-
Ef-B is a membrane-bound subunit, while subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex, Ef-AHD, which is also associated with the membrane
Manually annotated by BRENDA team
-
membrane-bound oxaloacetate decarboxylase complex, the alpha-subunit is a peripheral membrane protein on the cytosolic side of the membrane, where it associates with beta- and gamma-subunits that are embedded in the membrane. The beta-subunit is an integral membrane protein with nine transmembrane segments. The small gamma-subunit is an integral membrane protein with a single membrane-spanning helix at the N-terminus
Manually annotated by BRENDA team
-
Ef-B is a membrane-bound subunit, while subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex, Ef-AHD, which is also associated with the membrane
-
Manually annotated by BRENDA team
-
differs from cytoplasmic enzyme
Manually annotated by BRENDA team
-
14% of activity
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas putida (strain F1 / ATCC 700007)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
32000
-
-
monomer, SDS-PAGE
58000
-
-
gel filtration
62100
-
-
native enzyme, gel filtration
63000
-
-
gel filtration
63600
-
-
alpha-subunit, calculated from amino acid sequence
80000
-
-
gel filtration
98300
-
-
static light scattering
102000
-
-
gel filtration
105000
-
-
gel filtration
118000
-
-
gel filtration
530000
-
-
the most prominent band at 530 kDa is likely composed of a tetrameric Oad-alpha/gamma plus a dimeric (or tetrameric) Oad-beta subunit, SDS-PAGE
570000
-
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
oxaloacetate decarboxylase OAD-2, gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
2 * 49000, SDS-PAGE
dimer
-, Q9HUU1
dimer of dimers, X-ray crystallography
dimer
-
in the absence of beta- or alpha-subunits, the gamma-subunit forms a homodimer through a dimerization interface in the carboxyltransferase domain
heterotrimer
-
OAD contains three different subunits: Oad-alpha, a biotinylated extrinsic protein that catalyzes the alpha-ketodecarboxylation of oxaloacetate; Oad-gamma, a structural bitopic membrane protein whose cytosolic tail (named as Oad-gamma) binds tightly to Oad-alpha, and Oad-beta, a multispan transmembrane-alpha-helical protein that constitutes the Na+-channel
heterotrimer
-
-
homodimer
-
2 * 51000, gel filtration
homodimer
-
2 * 29000, gel filtration
tetramer
-
4 * 31700, SDS-PAGE
tetramer
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
oxaloacetate decarboxylase OAD-2
tetramer
-
the enzyme is constituted of four subunits: catalytic subunit OadA (termed Ef-A), membrane pump Ef-B, biotin acceptor protein Ef-D, and subunit Ef-H. Subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex, Ef-AHD, which is also associated with the membrane. The Ef-H subunit is involved in the cytoplasmic Ef-AHD complex formation. Analysis of subunit interactions and complex formation, overview
tetramer
-
the enzyme consists of alpha-, beta-, and gamma-subunits as well as a biotin carboxyl carrier protein domain. The 65 kDa hydrophilic alpha-subunit consists of an N-terminal carboxyltransferase domain connected to a C-terminal biotin carboxyl carrier protein domain. The 45 kDa beta-subunit is an integral membrane protein with nine transmembrane segments, which serves to couple the decarboxylation of carboxybiotin to the translocation of Na+ from the cytoplasm to the periplasm. The small 9 kDa gamma-subunit is an integral membrane protein with a single membrane-spanning helix at the N-terminus, followed by a hydrophilic C-terminal domain which interacts with the alpha-subunit. The gamma-subunit is essential for the overall stability of the complex, and likely serves as an anchor to hold the alpha- and beta-subunits in place
tetramer
-
the enzyme consists of alpha-, beta-, and gamma-subunits as well as a biotin carboxyl carrier protein domain. The about 65 kDa hydrophilic alpha-subunit consists of an N-terminal carboxyltransferase domain connected to a C-terminal biotin carboxyl carrier protein domain. The about 45 kDa beta-subunit is an integral membrane protein with nine transmembrane segments, which serves to couple the decarboxylation of carboxybiotin to the translocation of Na+ from the cytoplasm to the periplasm. the site of interaction with the gamma-subunit in Vibrio cholerae OADC is located in an intervening region between the carboxyltransferase and biotin carboxyl carrier protein domains of the alpha-subunit, termed the association domain. Tetramerization of alpha-OADC is mediated by an interaction between the association domain of the alpha-subunit and the cytosolic portion of the gamma-subunit in a manner. A biotin binding pocket, termed the exo-binding site, is located at the interface between the association domain and the carboxyltransferase domain. The interaction is facilitated by the tetramerization of alpha-OADC through interactions between the association domain and the the cytosolic portion of the gamma-subunit, which maintain two of the four alpha-OADC molecules in close proximity to the membrane-bound beta-subunit
tetramer
-
the enzyme is constituted of four subunits: catalytic subunit OadA (termed Ef-A), membrane pump Ef-B, biotin acceptor protein Ef-D, and subunit Ef-H. Subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex, Ef-AHD, which is also associated with the membrane. The Ef-H subunit is involved in the cytoplasmic Ef-AHD complex formation. Analysis of subunit interactions and complex formation, overview
-
tetramer
Vibrio cholerae O395-N1
-
oxaloacetate decarboxylase OAD-2
-
trimer
-
1 * 63600 + 1 * 34000 + 1 * 12000 alpha,beta,gamma, SDS-PAGE
trimer
-
1 * 65000 + 1 * 34000 + 1 * 12000 alpha,beta,gamma, SDS-PAGE, subunits carry different functions
trimer
-
alpha-subunit also necessary for Na+ transport, spatial arrangement and subunit interactions
trimer
-
spatial arrangement of cloned enzyme in Escherichia coli
trimer
-
1 * 63800 + 1 * 34500 + 1 * 10600, alpha,beta,gamma, SDS-PAGE
trimer
-
size and ultrastructure
trimer
-
alpha, beta, gamma
monomer
-
1 * 63000, SDS-PAGE
additional information
-
the alpha-subunit binds the gamma-subunit with a distinct association domain which is flanked on both sides with proline- and alanine-rich linker peptides
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapor diffusion method, using 0.2 M MgCl2, 0.1 M Bis-Tris pH 6.0, 25% (w/v) polyethylene glycol 3350
-
hanging drop vapour diffusion method, aliquots of 10 mg/ml protein in 5 mM MgCl2, 5 mM phosphonopyruvate and 10 mM NaHEPES (pH 7.0) mixed with equal volumes of reservoir solution containing 12% polyethylene glycol 20000 and 0.1 M MES (pH 6.0)
-, Q9HUU1
sitting drop vapour diffusion method using 0.1 M sodium cacodylate (pH 6.5), 0.2 M (NH4)2SO4, 5% (v/v) glycerol and 25% (w/v) PEG 8000
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
50
-
-
only if protected by oxalate
55
-
-
up to
85
-
-
half life for inacivation, 5 h
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
very instable, protease-sensitive, 50% loss of activity after 4 min at 4C
-
buffer-dependent, mercaptoethanol stabilises
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, Tris-HCl buffer, pH 7.5, 2 month, 40% loss of activity
-
frozen, Tris-HCl buffer, pH 7.5, 1 month, 25% loss of activity
-
-20C, potassium phosphate buffer, pH 7, 50% glycerol, 2 month
-
4C, glycerol, 2 months
Gluconacetobacter xylinus
-
25C, 50 mM triethanolamine/HCl, pH 7.0, 5 days, almost complete inactivation, presence of 1 mM Mn2+ and 50% v/v glycerol stabilize
-
4C, phosphate buffer or arsenate buffer, pH 7, with mercaptoethanol, N2-atmosphere, 2 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate precipitation, HiPrep column chromatography, Mono Q column chromatography, and Superdex-200 gel filtration
-
Ni-NTA column chromatography and Superdex G-200 gel filtration
-
alpha- and beta-chain
-
Ni2+-bounded affinity column chromatography
Q82YW6
recombinnat maltose-binding protein fusion protein OadH from Escherichia coli by amylase affinity chromatography, recombinant His-tagged subunits from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
-
Gluconacetobacter xylinus
-
recombinant enzyme from Escherichia coli
-
recombinant protein from Escherichia coli
-
recombinant
-
DEAE-cellulose column chromatography, phenyl-Sepharose columnn chromatography, and butyl-Sepharose column chromatography
-, Q9HUU1
avidin-Sepharose column chromatography
-
enzyme from Vibrio cholerae and heterologously expressed enzyme
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
Ni2+-NTA-agarose column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
expressed in Escherichia coli C43 (DE3) cells
-
expressed in Escherichia coli BL21(DE3) cells
Q82YW6
expression of gene oadH as maltose-binding protein fusion protein in Escherichia coli, genes oadA-oadD, expression of recombinant His-tagged subunits in Escherichia coli strain BL21(DE3)
-
all subunits, expressed in Escherichia coli, alpha-subunit strongly overexpressed from pT7-7 plasmid
-
alpha-subunit, expressed in Escherichia coli
-
expressed in Escherichia coli
-
expressed in Escherichia coli CC118
-
expressed in Escherichia coli DH5alpha
-
expressed in Escherichia coli DH5alpha/pSK-GAB
-
expression in Escherichia coli, expression of mutant enzymes and domains of the alpha subunit
-
expression in Escherichia coli M15
-
expressed in Escherichia coli BL21 (DE3) cells
-, Q9HUU1
expressed in Escherichia coli C43(DE3) cells
-
expressed in Escherichia coli Rosetta (C43) cells
-
expressed in Escherichia coli strain C43(DE3)
-
expression in Escherichia coli; expression in Escherichia coli
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
when subunit alpha-2 of oxaloacetate decarboxylase OAD-2 and the C-terminal domain of gamma-2 are synthesized together in Escherichia coli they form a complex that is stable at neutral pH and dissociates at pH-values below 5.0
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Y235A
-
site-directed mutagenesis
Y235F
-
site-directed mutagenesis
Y235S
-
site-directed mutagenesis
A383C
-
beta subunit, very low activity, low activity after modification with 2-aminoethyl methanethiosulfonate
C291E
-
beta subunit, unstable in the absence of Na+, dissociates into an alpha-gamma subcomplex and the beta subunit
C87A
-
slightly increased activity
C87A/A194C
-
no effect on activity
C87A/I198C
-
no effect on activity
C87A/L178C
-
about 50% of activity
C87A/L7C
-
no activity
C87A/P191C
-
no activity
C87A/P205C
-
no effect on activity; only traces of activity
C87A/S187C
-
no effect on activity
C87A/V129C
-
strongly increased activity
C87A/Y182C
-
slightly increased activity
D203N
-
beta subunit, complete loss of activity
D62A
-
gamma subunit, Zn2+ content decreased to 35% of the wild type enzyme, roughly 30% of activity
D63A
-
gamma subunit, almost no effect on activity
DELTAH76-P83
-
gamma subunit, unable to associate to the alpha subunit, no activity
DELTAH78-P83
-
gamma subunit, unable to associate to the alpha subunit, no activity
DELTAH82-P83
-
gamma subunit, Zn2+ content decreased to 5% of the wild type enzyme, very low activity
G337A
-
beta subunit, complete loss of activity
G377A
-
beta subunit, complete loss of activity
G377C
-
beta subunit, no activity even after modification with 2-aminoethyl methanethiosulfonate
G380A
-
beta subunit, about 30% of activity
G380C
-
beta subunit, very low activity, low activity after modification with 2-aminoethyl methanethiosulfonate
H76A
-
gamma subunit, almost no effect on activity
H77A
-
gamma subunit, Zn2+ content decreased to 10% of the wild type enzyme, very low activity
N373D
-
beta subunit, no protection against tryptic digestion by Na+
N373L
-
beta subunit, almost complete loss of activity
N373L
-
beta subunit, very low activity, no protection against tryptic digestion by Na+
N392L
-
beta subunit, about 70% of activity
Q192L
-
beta subunit, lower pH optimum than wild type enzyme
R389A
-
beta subunit, about 10% of activity, significantly decreased Na+ binding
R389A
-
beta subunit, significantly increased pH-optimum
R389C
-
beta subunit, increased pH-optimum, reduced activity, activity and pH-optimum can be restored to wild type value by modification with 2-aminoethyl methanethiosulfonate
R389D
-
beta subunit, almost complete loss of activity
R389D
-
beta subunit, very low activity
R389K
-
beta subunit, about 80% of activity, significantly decreased Na+ binding
R389L
-
beta subunit, about 10% of activity, significantly decreased Na+ binding
S382A
-
beta subunit, complete loss of activity
S382A
-
beta subunit, no activity below 20 mM Na+, active with 400 mM NaCl, no protection against tryptic digestion by Na+, no Na+ transport
S382C
-
beta subunit, complete loss of activity
S382C
-
beta subunit, no activity even after modification with 2-aminoethyl methanethiosulfonate
S382D
-
beta subunit, about 10% of activity
S382D
-
beta subunit, very low activity
S382E
-
beta subunit, complete loss of activity
S382N
-
beta subunit, complete loss of activity
S382Q
-
beta subunit, complete loss of activity
S382T
-
beta subunit, about 10% of activity
S382T
-
beta subunit, very low activity
Y209A
-
beta subunit, lower pH optimum than wild type enzyme
H235A
-, Q9HUU1
the kcat value for catalysis of oxaloacetate decarboxylation is less than an order of magnitude smaller compared to the wild type enzyme
H235Q
-, Q9HUU1
the kcat value for catalysis of oxaloacetate decarboxylation is less than an order of magnitude smaller compared to the wild type enzyme
Y212F
-, Q9HUU1
25fold reduction of the Km value compared to the wild type enzyme
C148A
-
74% activity of the wild type enzyme
C148S
-
54% activity of the wild type enzyme
D17A
-
inactive
H207I
-
inactive
H209I
-
inactive
K178I
-
inactive
K178R
-
inactive
M180I
-
16% activity of the wild type enzyme
Q20L
-
inactive
R16I
-
inactive
additional information
-
oadA, oadB, and oadD deletion mutants are constructed, while an oadH mutant strain cannot be obtained in spite of using different genetic strategies
additional information
-
oadA, oadB, and oadD deletion mutants are constructed, while an oadH mutant strain cannot be obtained in spite of using different genetic strategies
-
H78A
-
gamma subunit, binding to alpha subunit is lost; gamma subunit, unable to associate to the alpha subunit, no activity
additional information
-
fusion protein from different domains of the enzyme subunits
additional information
-
mutations in the beta subunit do not affect the ability to transfer the carboxyl-group to biotin, but reduce the decarboxylase activity
additional information
-
several other mutant with reduced activity
Y229F
-
beta subunit, completely inactive
additional information
-
mutant [ILCitM (pFL3)] is constructed by double homologous recombination, during culture with citrate the growth rate of the mutant is lower than the willd type enzyme
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
reconstition of isolated alpha-chains and bacterial membranes containing beta- and gamma-chains
-
heterologously expressed oxaloacetate decarboxylase OAD-2
Q6A1F5, Q6A1F6, Q6A1F7, Q6A1G0, Q9KUH0, Q9KUH1, -
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
optimization of oxaloacetate may increase lysine productivity of commercially used strains