Biosynthesis of vitamin B12: stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide is catalyzed by one enzyme in Pseudomonas denitrificans
Blanche, F.; Couder, M.; Debussche, L.; Thibaut, D.; Cameron, B.; Cruzet, J.; J. Bacteriol. 173, 6046-6051 (1991) 
Data extracted from this reference:
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
6.3.5.9
-
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
cobQ gene
Pseudomonas denitrificans (nom. rej.)
General Stability
EC Number
General Stability
Organism
6.3.5.9
glycerol and DTT are necessary to stabilize the enzyme during purification
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
20% w/v and 1 mM dithiothreitol are necessary to stabilize enzyme during purification
Pseudomonas denitrificans (nom. rej.)
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.3.5.9
0.00104
-
adenosyl-cobyrinic acid a,c-diamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.00121
-
adenosyl-cobyrinic acid tetraamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.00122
-
adenosyl-cobyrinic acid triamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.00125
-
adenosyl-cobyrinic acid pentaamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.011
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.033
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.035
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.045
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.049
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.064
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.088
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.12
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
20
-
ammonia
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
additional information
-
additional information
-
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00104
-
adenosylcobyrinic acid a,c-diamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00121
-
adenosylcobyrinic acid tetraamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00122
-
adenosylcobyrinic acid triamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00125
-
adenosylcobyrinic acid pentaamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.011
-
ATP
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.033
-
L-glutamine
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.035
-
ATP
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.045
-
L-glutamine
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.049
-
L-glutamine
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.064
-
L-glutamine
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.088
-
ATP
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.12
-
ATP
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
20.4
-
ammonia
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.5.9
Mg2+
amidation reaction is Mg2+ dependent
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
Mg2+
Mg2+-dependent
Pseudomonas denitrificans (nom. rej.)
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.3.5.9
57000
-
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
97300
-
gel filtration
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
52000
-
predicted from DNA sequence of the cobQ gene
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
57000
-
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
97300
-
gel filtration
Pseudomonas denitrificans (nom. rej.)
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
Pseudomonas denitrificans (nom. rej.)
cobalamin biosynthetic pathway
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
Pseudomonas denitrificans (nom. rej.)
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
Pseudomonas denitrificans (nom. rej.) SC510 RifT
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
6.3.5.9
Pseudomonas denitrificans (nom. rej.)
-
-
-
6.3.5.9
Pseudomonas denitrificans (nom. rej.) SC510 RifT
-
-
-
6.3.5.10
Pseudomonas denitrificans (nom. rej.)
-
-
-
6.3.5.10
Pseudomonas denitrificans (nom. rej.)
-
recombinant strain SC510 Rifr(pXL617)
-
Oxidation Stability
EC Number
Oxidation Stability
Organism
6.3.5.10
sensitive to oxidation
Pseudomonas denitrificans (nom. rej.)
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
6.3.5.9
-
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
45fold
Pseudomonas denitrificans (nom. rej.)
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
6.3.5.9
0.068
-
adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.083
-
adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.083
-
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
Storage Stability
EC Number
Storage Stability
Organism
6.3.5.9
-20°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
-20°C, enzyme can be stored in the eluate from purification step v without detectable loss of activity
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
-20°C or 0°C, purified enzyme, 0.1 M Tris hydrochloride, pH 7.5, 48 h, 100% loss of activity
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
-20°C, purified enzyme, 50 mM Tris hydrochloride, pH 7.5, 0.1 M sodium chloride, 1 mM dithiothreitol, 20% glycerol, several weeks, stable
Pseudomonas denitrificans (nom. rej.)
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nom. rej.)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + ATP + ammonia + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + ADP + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
adenosyl-cobyrinic acid pentaamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
adenosyl-cobyrinic acid tetraamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
adenosyl-cobyrinic acid triamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
ATP + adenosyl-cobyrinic acid a,c-diamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + ?
-
-
-
?
6.3.5.9
additional information
adenosyl-cobyric acid and hydrogenobyrinic acid a,c-diamide are no substrates
651618
Pseudomonas denitrificans (nom. rej.)
?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + ammonia + H2O
ammonia can replace L-glutamine as amide group donor, but L-glutamine is preferred
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + ?
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
enzyme catalyzes the stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide, four-step amidation sequence from cobyrinic acid a,c-diamide to cobyric acid via the formation of corbyrinic acid triamide, tetraamide, and pentaamide intermediates, the amidations are carried out in a specific order, enzyme is specific to coenzyme forms of substrates, no amidation of the carboxyl group at position f, L-glutamine is the preferred amide group donor
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
enzyme catalyzes the stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide, four-step amidation sequence from cobyrinic acid a,c-diamide to cobyric acid via the formation of corbyrinic acid triamide, tetraamide, and pentaamide intermediates, the amidations are carried out in a specific order, enzyme is specific to coenzyme forms of substrates, no amidation of the carboxyl group at position f, L-glutamine is the preferred amide group donor
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid pentaamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid tetraamide + L-glutamine + H2O
two-step amidation
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid triamide + L-glutamine + H2O
three-step amidation
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
?
6.3.5.10
additional information
no amidation of adenosylcobyric acid, hydrogenobyrinic acid a,c-diamide, monocyano or diaqua form of cobyrinic acid a,c-diamide
651618
Pseudomonas denitrificans (nom. rej.)
?
-
-
-
?
6.3.5.10
additional information
no amidation of adenosylcobyric acid, hydrogenobyrinic acid a,c-diamide, monocyano or diaqua form of cobyrinic acid a,c-diamide
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
6.3.5.9
dimer
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
dimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
homodimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
Synonyms
EC Number
Synonyms
Commentary
Organism
6.3.5.9
cobyric acid synthase
-
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
CobQ
-
Pseudomonas denitrificans (nom. rej.)
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.3.5.9
7.5
-
broad optimum around
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
7.5
-
broad pH-optimum around
Pseudomonas denitrificans (nom. rej.)
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
6.3.5.10
ATP
ATP-dependent
Pseudomonas denitrificans (nom. rej.)
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
6.3.5.9
Pseudomonas denitrificans (nom. rej.)
-
5.9
5.6
6.3.5.10
Pseudomonas denitrificans (nom. rej.)
isoelectric focusing
-
6
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
6.3.5.9
-
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
cobQ gene
Pseudomonas denitrificans (nom. rej.)
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
6.3.5.10
ATP
ATP-dependent
Pseudomonas denitrificans (nom. rej.)
General Stability (protein specific)
EC Number
General Stability
Organism
6.3.5.9
glycerol and DTT are necessary to stabilize the enzyme during purification
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
20% w/v and 1 mM dithiothreitol are necessary to stabilize enzyme during purification
Pseudomonas denitrificans (nom. rej.)
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.3.5.9
0.00104
-
adenosyl-cobyrinic acid a,c-diamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.00121
-
adenosyl-cobyrinic acid tetraamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.00122
-
adenosyl-cobyrinic acid triamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.00125
-
adenosyl-cobyrinic acid pentaamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.011
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.033
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.035
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.045
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.049
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.064
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.088
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.12
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
20
-
ammonia
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
additional information
-
additional information
-
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00104
-
adenosylcobyrinic acid a,c-diamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00121
-
adenosylcobyrinic acid tetraamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00122
-
adenosylcobyrinic acid triamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.00125
-
adenosylcobyrinic acid pentaamide
pH 7.5
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.011
-
ATP
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.033
-
L-glutamine
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.035
-
ATP
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.045
-
L-glutamine
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.049
-
L-glutamine
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.064
-
L-glutamine
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.088
-
ATP
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.12
-
ATP
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
20.4
-
ammonia
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.5.9
Mg2+
amidation reaction is Mg2+ dependent
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
Mg2+
Mg2+-dependent
Pseudomonas denitrificans (nom. rej.)
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.3.5.9
57000
-
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
97300
-
gel filtration
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
52000
-
predicted from DNA sequence of the cobQ gene
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
57000
-
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
97300
-
gel filtration
Pseudomonas denitrificans (nom. rej.)
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
Pseudomonas denitrificans (nom. rej.)
cobalamin biosynthetic pathway
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
Pseudomonas denitrificans (nom. rej.)
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
Pseudomonas denitrificans (nom. rej.) SC510 RifT
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
?
Oxidation Stability (protein specific)
EC Number
Oxidation Stability
Organism
6.3.5.10
sensitive to oxidation
Pseudomonas denitrificans (nom. rej.)
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
6.3.5.9
-
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
45fold
Pseudomonas denitrificans (nom. rej.)
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
6.3.5.9
0.068
-
adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0.083
-
adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
0.083
-
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
6.3.5.9
-20°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
-20°C, enzyme can be stored in the eluate from purification step v without detectable loss of activity
Pseudomonas denitrificans (nom. rej.)
6.3.5.9
0°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
-20°C or 0°C, purified enzyme, 0.1 M Tris hydrochloride, pH 7.5, 48 h, 100% loss of activity
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
-20°C, purified enzyme, 50 mM Tris hydrochloride, pH 7.5, 0.1 M sodium chloride, 1 mM dithiothreitol, 20% glycerol, several weeks, stable
Pseudomonas denitrificans (nom. rej.)
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nom. rej.)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + ATP + ammonia + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + ADP + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
adenosyl-cobyrinic acid pentaamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
adenosyl-cobyrinic acid tetraamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
adenosyl-cobyrinic acid triamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.9
ATP + adenosyl-cobyrinic acid a,c-diamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + ?
-
-
-
?
6.3.5.9
additional information
adenosyl-cobyric acid and hydrogenobyrinic acid a,c-diamide are no substrates
651618
Pseudomonas denitrificans (nom. rej.)
?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + ammonia + H2O
ammonia can replace L-glutamine as amide group donor, but L-glutamine is preferred
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + ?
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
enzyme catalyzes the stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide, four-step amidation sequence from cobyrinic acid a,c-diamide to cobyric acid via the formation of corbyrinic acid triamide, tetraamide, and pentaamide intermediates, the amidations are carried out in a specific order, enzyme is specific to coenzyme forms of substrates, no amidation of the carboxyl group at position f, L-glutamine is the preferred amide group donor
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
enzyme catalyzes the stepwise amidation of carboxyl groups b, d, e, and g of cobyrinic acid a,c-diamide, four-step amidation sequence from cobyrinic acid a,c-diamide to cobyric acid via the formation of corbyrinic acid triamide, tetraamide, and pentaamide intermediates, the amidations are carried out in a specific order, enzyme is specific to coenzyme forms of substrates, no amidation of the carboxyl group at position f, L-glutamine is the preferred amide group donor
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid pentaamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid tetraamide + L-glutamine + H2O
two-step amidation
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid triamide + L-glutamine + H2O
three-step amidation
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.)
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
?
6.3.5.10
additional information
no amidation of adenosylcobyric acid, hydrogenobyrinic acid a,c-diamide, monocyano or diaqua form of cobyrinic acid a,c-diamide
651618
Pseudomonas denitrificans (nom. rej.)
?
-
-
-
?
6.3.5.10
additional information
no amidation of adenosylcobyric acid, hydrogenobyrinic acid a,c-diamide, monocyano or diaqua form of cobyrinic acid a,c-diamide
651618
Pseudomonas denitrificans (nom. rej.) SC510 RifT
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
6.3.5.9
dimer
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
dimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
homodimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.3.5.9
7.5
-
broad optimum around
Pseudomonas denitrificans (nom. rej.)
6.3.5.10
7.5
-
broad pH-optimum around
Pseudomonas denitrificans (nom. rej.)
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
6.3.5.9
Pseudomonas denitrificans (nom. rej.)
-
5.9
5.6
6.3.5.10
Pseudomonas denitrificans (nom. rej.)
isoelectric focusing
-
6