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
Commentary
Organism
6.3.5.9
-
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
cobQ gene
Pseudomonas denitrificans (nomen rejiciendum)
General Stability
EC Number
General Stability
Organism
6.3.5.9
glycerol and DTT are necessary to stabilize the enzyme during purification
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
20% w/v and 1 mM dithiothreitol are necessary to stabilize enzyme during purification
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
0.00121
-
adenosyl-cobyrinic acid tetraamide
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.00122
-
adenosyl-cobyrinic acid triamide
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.00125
-
adenosyl-cobyrinic acid pentaamide
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.011
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.033
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.035
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.045
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.049
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.064
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.088
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.12
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
20
-
ammonia
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
additional information
-
additional information
-
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00104
-
adenosylcobyrinic acid a,c-diamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00121
-
adenosylcobyrinic acid tetraamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00122
-
adenosylcobyrinic acid triamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00125
-
adenosylcobyrinic acid pentaamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.011
-
ATP
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.033
-
L-glutamine
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.035
-
ATP
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.045
-
L-glutamine
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.049
-
L-glutamine
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.064
-
L-glutamine
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.088
-
ATP
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.12
-
ATP
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
20.4
-
ammonia
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.5.9
Mg2+
amidation reaction is Mg2+ dependent
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
Mg2+
Mg2+-dependent
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
97300
-
gel filtration
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
52000
-
predicted from DNA sequence of the cobQ gene
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
57000
-
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
97300
-
gel filtration
Pseudomonas denitrificans (nomen rejiciendum)
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
Pseudomonas denitrificans (nomen rejiciendum)
cobalamin biosynthetic pathway
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum) SC510 RifT
biosynthesis of vitamin B12, cobalamin biosynthetic pathway
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
6.3.5.9
Pseudomonas denitrificans (nomen rejiciendum)
-
-
-
6.3.5.9
Pseudomonas denitrificans (nomen rejiciendum) SC510 RifT
-
-
-
6.3.5.10
Pseudomonas denitrificans (nomen rejiciendum)
-
; recombinant strain SC510 Rifr(pXL617)
-
Oxidation Stability
EC Number
Oxidation Stability
Organism
6.3.5.10
sensitive to oxidation
Pseudomonas denitrificans (nomen rejiciendum)
Purification (Commentary)
EC Number
Commentary
Organism
6.3.5.9
-
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
45fold
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
0.083
-
adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.083
-
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
-20°C, enzyme can be stored in the eluate from purification step v without detectable loss of activity
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
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 (nomen rejiciendum)
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nomen rejiciendum)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + ATP + ammonia + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + ADP + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
adenosyl-cobyrinic acid pentaamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
adenosyl-cobyrinic acid tetraamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
adenosyl-cobyrinic acid triamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
additional information
adenosyl-cobyric acid and hydrogenobyrinic acid a,c-diamide are no substrates
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum) SC510 RifT
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum) 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 (nomen rejiciendum)
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 (nomen rejiciendum)
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 (nomen rejiciendum)
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 (nomen rejiciendum) 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 (nomen rejiciendum) SC510 RifT
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid pentaamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid tetraamide + L-glutamine + H2O
two-step amidation
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid triamide + L-glutamine + H2O
three-step amidation
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum) SC510 RifT
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum) 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 (nomen rejiciendum)
?
-
-
-
-
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 (nomen rejiciendum) SC510 RifT
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
6.3.5.9
dimer
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
dimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
homodimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.3.5.9
7.5
-
broad optimum around
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
7.5
-
broad pH-optimum around
Pseudomonas denitrificans (nomen rejiciendum)
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
6.3.5.10
ATP
ATP-dependent
Pseudomonas denitrificans (nomen rejiciendum)
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
6.3.5.9
Pseudomonas denitrificans (nomen rejiciendum)
-
5.9
5.6
6.3.5.10
Pseudomonas denitrificans (nomen rejiciendum)
isoelectric focusing
-
6
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
6.3.5.9
-
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
cobQ gene
Pseudomonas denitrificans (nomen rejiciendum)
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
6.3.5.10
ATP
ATP-dependent
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.10
20% w/v and 1 mM dithiothreitol are necessary to stabilize enzyme during purification
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
0.00121
-
adenosyl-cobyrinic acid tetraamide
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.00122
-
adenosyl-cobyrinic acid triamide
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.00125
-
adenosyl-cobyrinic acid pentaamide
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.011
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.033
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.035
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.045
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.049
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.064
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.088
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0.12
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
20
-
ammonia
pH 7.5, 20°C
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
additional information
-
additional information
-
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00104
-
adenosylcobyrinic acid a,c-diamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00121
-
adenosylcobyrinic acid tetraamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00122
-
adenosylcobyrinic acid triamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.00125
-
adenosylcobyrinic acid pentaamide
pH 7.5
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.011
-
ATP
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.033
-
L-glutamine
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.035
-
ATP
pH 7.5, adenosylcobyrinic acid triamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.045
-
L-glutamine
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.049
-
L-glutamine
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.064
-
L-glutamine
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.088
-
ATP
pH 7.5, adenosylcobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.12
-
ATP
pH 7.5, adenosylcobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
20.4
-
ammonia
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.5.9
Mg2+
amidation reaction is Mg2+ dependent
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
Mg2+
Mg2+-dependent
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
97300
-
gel filtration
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
52000
-
predicted from DNA sequence of the cobQ gene
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
57000
-
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
97300
-
gel filtration
Pseudomonas denitrificans (nomen rejiciendum)
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
Pseudomonas denitrificans (nomen rejiciendum)
cobalamin biosynthetic pathway
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum) 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 (nomen rejiciendum)
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
6.3.5.9
-
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
45fold
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
0.083
-
adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
0.083
-
pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.9
-20°C, enzyme can be stored in the eluate from purification step v without detectable loss of activity
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.9
0°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
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 (nomen rejiciendum)
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
cobalamin biosynthetic pathway
651618
Pseudomonas denitrificans (nomen rejiciendum)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
5'-deoxy-5'-adenosyl-cobyrinic acid + ATP + ammonia + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + ADP + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
adenosyl-cobyrinic acid pentaamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
adenosyl-cobyrinic acid tetraamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
adenosyl-cobyrinic acid triamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.9
additional information
adenosyl-cobyric acid and hydrogenobyrinic acid a,c-diamide are no substrates
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum) SC510 RifT
adenosylcobyric acid + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum) 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 (nomen rejiciendum)
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 (nomen rejiciendum)
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 (nomen rejiciendum)
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 (nomen rejiciendum) 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 (nomen rejiciendum) SC510 RifT
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid pentaamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid tetraamide + L-glutamine + H2O
two-step amidation
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
ATP + adenosylcobyrinic acid triamide + L-glutamine + H2O
three-step amidation
651618
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + adenosylcobyric acid + L-glutamate
-
-
-
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum)
?
6.3.5.10
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nomen rejiciendum) SC510 RifT
cobinamide + ADP + phosphate + L-glutamate
-
651618
Pseudomonas denitrificans (nomen rejiciendum) 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 (nomen rejiciendum)
?
-
-
-
-
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 (nomen rejiciendum) SC510 RifT
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
6.3.5.9
dimer
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
dimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.3.5.10
homodimer
2 * 57000, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
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 (nomen rejiciendum)
6.3.5.10
7.5
-
broad pH-optimum around
Pseudomonas denitrificans (nomen rejiciendum)
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
6.3.5.9
Pseudomonas denitrificans (nomen rejiciendum)
-
5.9
5.6
6.3.5.10
Pseudomonas denitrificans (nomen rejiciendum)
isoelectric focusing
-
6