BRENDA - Enzyme Database
show all sequences of 6.3.5.9

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)
Cloned (Commentary)
Organism
-
Pseudomonas denitrificans (nom. rej.)
General Stability
General Stability
Organism
glycerol and DTT are necessary to stabilize the enzyme during purification
Pseudomonas denitrificans (nom. rej.)
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00104
-
adenosyl-cobyrinic acid a,c-diamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.00121
-
adenosyl-cobyrinic acid tetraamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.00122
-
adenosyl-cobyrinic acid triamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.00125
-
adenosyl-cobyrinic acid pentaamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.011
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.033
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.035
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.045
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.049
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.064
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.088
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.12
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
20
-
ammonia
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
amidation reaction is Mg2+ dependent
Pseudomonas denitrificans (nom. rej.)
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
57000
-
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
97300
-
gel filtration
Pseudomonas denitrificans (nom. rej.)
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
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.)
?
Organism
Organism
UniProt
Commentary
Textmining
Pseudomonas denitrificans (nom. rej.)
-
-
-
Pseudomonas denitrificans (nom. rej.) SC510 RifT
-
-
-
Purification (Commentary)
Purification (Commentary)
Organism
-
Pseudomonas denitrificans (nom. rej.)
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.068
-
adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.083
-
adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
Storage Stability
Storage Stability
Organism
-20°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
-20°C, enzyme can be stored in the eluate from purification step v without detectable loss of activity
Pseudomonas denitrificans (nom. rej.)
0°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
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.)
?
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.)
?
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.)
?
adenosyl-cobyrinic acid pentaamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
adenosyl-cobyrinic acid tetraamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
adenosyl-cobyrinic acid triamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
ATP + adenosyl-cobyrinic acid a,c-diamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + ?
-
-
-
?
additional information
adenosyl-cobyric acid and hydrogenobyrinic acid a,c-diamide are no substrates
651618
Pseudomonas denitrificans (nom. rej.)
?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
Subunits
Subunits
Commentary
Organism
dimer
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
Synonyms
Synonyms
Commentary
Organism
cobyric acid synthase
-
Pseudomonas denitrificans (nom. rej.)
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
broad optimum around
Pseudomonas denitrificans (nom. rej.)
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Pseudomonas denitrificans (nom. rej.)
-
5.9
5.6
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Pseudomonas denitrificans (nom. rej.)
General Stability (protein specific)
General Stability
Organism
glycerol and DTT are necessary to stabilize the enzyme during purification
Pseudomonas denitrificans (nom. rej.)
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00104
-
adenosyl-cobyrinic acid a,c-diamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.00121
-
adenosyl-cobyrinic acid tetraamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.00122
-
adenosyl-cobyrinic acid triamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.00125
-
adenosyl-cobyrinic acid pentaamide
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
0.011
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.033
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.035
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid triamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.045
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.049
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.064
-
L-glutamine
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.088
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.12
-
ATP
pH 7.5, 20°C, adenosyl-cobyrinic acid tetraamide as substrate
Pseudomonas denitrificans (nom. rej.)
20
-
ammonia
pH 7.5, 20°C
Pseudomonas denitrificans (nom. rej.)
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
amidation reaction is Mg2+ dependent
Pseudomonas denitrificans (nom. rej.)
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
57000
-
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
97300
-
gel filtration
Pseudomonas denitrificans (nom. rej.)
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
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.)
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Pseudomonas denitrificans (nom. rej.)
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.068
-
adenosyl-cobyrinic acid a,c-diamide as substrate
Pseudomonas denitrificans (nom. rej.)
0.083
-
adenosyl-cobyrinic acid pentaamide as substrate
Pseudomonas denitrificans (nom. rej.)
Storage Stability (protein specific)
Storage Stability
Organism
-20°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
-20°C, enzyme can be stored in the eluate from purification step v without detectable loss of activity
Pseudomonas denitrificans (nom. rej.)
0°C, 0.1 M Tris hydrochloride, pH 7.5, 100% activity is lost after 48 h
Pseudomonas denitrificans (nom. rej.)
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
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.)
?
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.)
?
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.)
?
adenosyl-cobyrinic acid pentaamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
adenosyl-cobyrinic acid tetraamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
adenosyl-cobyrinic acid triamide + ATP + glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + glutamate + phosphate + ?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
ATP + adenosyl-cobyrinic acid a,c-diamide + L-glutamine + H2O
-
651618
Pseudomonas denitrificans (nom. rej.)
ADP + phosphate + ?
-
-
-
?
additional information
adenosyl-cobyric acid and hydrogenobyrinic acid a,c-diamide are no substrates
651618
Pseudomonas denitrificans (nom. rej.)
?
-
651618
Pseudomonas denitrificans (nom. rej.)
?
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
2 * 57000, homodimer, SDS-PAGE
Pseudomonas denitrificans (nom. rej.)
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
broad optimum around
Pseudomonas denitrificans (nom. rej.)
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Pseudomonas denitrificans (nom. rej.)
-
5.9
5.6
Other publictions for EC 6.3.5.9
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
661083
Fresquet
Mechanism of cobyrinic acid a, ...
Salmonella enterica subsp. enterica serovar Typhimurium
Biochemistry
43
10619-10627
2004
-
-
1
-
6
-
-
8
-
-
2
-
-
1
-
-
-
-
-
-
-
-
4
1
1
-
-
-
8
1
-
-
-
-
-
-
-
-
1
-
-
6
-
-
-
-
8
-
-
2
-
-
-
-
-
-
-
-
-
4
1
-
-
-
8
1
-
-
-
-
-
-
-
-
-
653832
Galperin
The synthetase domains of coba ...
Escherichia coli
Proteins
41
238-247
2000
-
-
-
-
-
-
-
-
-
-
1
1
-
4
-
-
-
-
-
-
-
-
2
1
3
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
2
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
651906
Tsang
CobB, a new member of the SIR2 ...
Salmonella enterica subsp. enterica serovar Typhimurium
J. Biol. Chem.
273
31788-31794
1998
-
-
1
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
651627
Tsang
cobB function is required for ...
Salmonella enterica subsp. enterica serovar Typhimurium
J. Bacteriol.
178
7016-7019
1996
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
3
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
651618
Blanche
Biosynthesis of vitamin B12: s ...
Pseudomonas denitrificans (nom. rej.), Pseudomonas denitrificans (nom. rej.) SC510 RifT
J. Bacteriol.
173
6046-6051
1991
-
-
1
-
-
1
-
13
-
1
2
1
-
2
-
-
1
-
-
-
2
3
8
1
1
-
-
-
-
1
-
-
-
-
1
-
-
-
1
-
-
-
1
-
-
-
13
-
1
2
1
-
-
-
1
-
-
2
3
8
1
-
-
-
-
1
-
-
1
-
-
-
-
-
-
485086
Crouzet
Nucleotide sequence of a Pseud ...
Pseudomonas denitrificans (nom. rej.), Pseudomonas denitrificans (nom. rej.) SC510
J. Bacteriol.
172
5968-5979
1990
-
-
1
-
-
-
-
-
-
-
1
2
-
4
-
-
1
-
-
-
-
-
6
1
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
1
2
-
-
-
1
-
-
-
-
6
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
644524
Debussche
Purification and characterizat ...
Pseudomonas denitrificans (nom. rej.)
J. Bacteriol.
172
6239-6244
1990
-
-
1
-
-
-
-
7
-
1
2
2
-
4
-
-
1
-
-
-
-
-
11
1
1
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
7
-
1
2
2
-
-
-
1
-
-
-
-
11
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-