BRENDA - Enzyme Database show
show all sequences of 6.6.1.2

Assay, purification, and characterization of cobaltochelatase, a unique complex enzyme catalyzing cobalt insertion in hydrogenobyrinic acid a,c-diamide during coenzime B12 biosynthesis in Pseudomonas denitrificans

Debussche, L.; Couder, M.; Thibaut, D.; Cameron, B.; Crouzet, J.; Blanche, F.; J. Bacteriol. 174, 7445-7451 (1992)

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
Organism
Structure
adenylyl (beta,gamma-methylene)-diphosphonate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
adenylyl-imidodiphosphate
50% inhibition at 2 mM
Pseudomonas denitrificans (nomen rejiciendum)
ADP
50% inhibition at 8 mM
Pseudomonas denitrificans (nomen rejiciendum)
AMP
15% inhibition at 10 mM
Pseudomonas denitrificans (nomen rejiciendum)
Cu2+
50% inhibition at 0.002 mM
Pseudomonas denitrificans (nomen rejiciendum)
diphosphate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
Fe2+
50% inhibition at 0.05 mM
Pseudomonas denitrificans (nomen rejiciendum)
N-ethylmaleimide
50% inhibition at 0.005 mM
Pseudomonas denitrificans (nomen rejiciendum)
Ni2+
50% inhibition at 0.003 mM
Pseudomonas denitrificans (nomen rejiciendum)
Zn2+
50% inhibition at 0.0005 mM
Pseudomonas denitrificans (nomen rejiciendum)
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.000085
-
hydrogenobyrinic acid a,c-diamide
pH 8.0, 30°C
Pseudomonas denitrificans (nomen rejiciendum)
0.0042
-
Co2+
pH 8.0, 30°C
Pseudomonas denitrificans (nomen rejiciendum)
0.22
-
ATP
pH 8.0, 30°C
Pseudomonas denitrificans (nomen rejiciendum)
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
ATP binding cation
Pseudomonas denitrificans (nomen rejiciendum)
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
80000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
140000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
258000
-
gel filtration
Pseudomonas denitrificans (nomen rejiciendum)
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
hydrogenobyrinic acid a,c-diamide + ATP + Co2+
Pseudomonas denitrificans (nomen rejiciendum)
-
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
Pseudomonas denitrificans (nomen rejiciendum)
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas denitrificans (nomen rejiciendum)
P29933 AND P29934 AND Q9HZQ3 AND P29929
-
-
Purification (Commentary)
Commentary
Organism
purification includes: Mono Q HR 10/10, Penyl-Superose and Mono Q HR 5/5 chromatographies for CobN subunit, Mono Q HR 10/10, Phenyl-Superose and AGATP for CobST subunit
Pseudomonas denitrificans (nomen rejiciendum)
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
0.0057 units for CobST subunit, 1 unit of CobN is defined as the amount of CobST necessary to reconstruct 1 unit of the entire complex. 0.006 for CobN subunit, 1 unit of CobN is defined as the amount of CobN necessary to reconstruct 1 unit of the entire complex
Pseudomonas denitrificans (nomen rejiciendum)
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + hydrogenobyrinic acid a,c-diamide + Co2+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
CTP + hydrogenobyrinic acid a,c-diamide + Co2+
45% of activity with respect to ATP
653078
Pseudomonas denitrificans (nomen rejiciendum)
CDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
dATP + hydrogenobyrinic acid a,c-diamide + Co2+
65% of activity with respect to ATP
653078
Pseudomonas denitrificans (nomen rejiciendum)
dADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
hydrogenobyrinic acid a,c-diamide + ATP + Co2+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
ITP + hydrogenobyrinic acid a,c-diamide + Co2+
5% of activity with respect to ATP
653078
Pseudomonas denitrificans (nomen rejiciendum)
IDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
Subunits
Subunits
Commentary
Organism
trimer
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
adenylyl (beta,gamma-methylene)-diphosphonate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
adenylyl-imidodiphosphate
50% inhibition at 2 mM
Pseudomonas denitrificans (nomen rejiciendum)
ADP
50% inhibition at 8 mM
Pseudomonas denitrificans (nomen rejiciendum)
AMP
15% inhibition at 10 mM
Pseudomonas denitrificans (nomen rejiciendum)
Cu2+
50% inhibition at 0.002 mM
Pseudomonas denitrificans (nomen rejiciendum)
diphosphate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
Fe2+
50% inhibition at 0.05 mM
Pseudomonas denitrificans (nomen rejiciendum)
N-ethylmaleimide
50% inhibition at 0.005 mM
Pseudomonas denitrificans (nomen rejiciendum)
Ni2+
50% inhibition at 0.003 mM
Pseudomonas denitrificans (nomen rejiciendum)
Zn2+
50% inhibition at 0.0005 mM
Pseudomonas denitrificans (nomen rejiciendum)
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.000085
-
hydrogenobyrinic acid a,c-diamide
pH 8.0, 30°C
Pseudomonas denitrificans (nomen rejiciendum)
0.0042
-
Co2+
pH 8.0, 30°C
Pseudomonas denitrificans (nomen rejiciendum)
0.22
-
ATP
pH 8.0, 30°C
Pseudomonas denitrificans (nomen rejiciendum)
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
ATP binding cation
Pseudomonas denitrificans (nomen rejiciendum)
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
80000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
140000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
258000
-
gel filtration
Pseudomonas denitrificans (nomen rejiciendum)
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
hydrogenobyrinic acid a,c-diamide + ATP + Co2+
Pseudomonas denitrificans (nomen rejiciendum)
-
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
Pseudomonas denitrificans (nomen rejiciendum)
?
Purification (Commentary) (protein specific)
Commentary
Organism
purification includes: Mono Q HR 10/10, Penyl-Superose and Mono Q HR 5/5 chromatographies for CobN subunit, Mono Q HR 10/10, Phenyl-Superose and AGATP for CobST subunit
Pseudomonas denitrificans (nomen rejiciendum)
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
0.0057 units for CobST subunit, 1 unit of CobN is defined as the amount of CobST necessary to reconstruct 1 unit of the entire complex. 0.006 for CobN subunit, 1 unit of CobN is defined as the amount of CobN necessary to reconstruct 1 unit of the entire complex
Pseudomonas denitrificans (nomen rejiciendum)
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + hydrogenobyrinic acid a,c-diamide + Co2+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
CTP + hydrogenobyrinic acid a,c-diamide + Co2+
45% of activity with respect to ATP
653078
Pseudomonas denitrificans (nomen rejiciendum)
CDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
dATP + hydrogenobyrinic acid a,c-diamide + Co2+
65% of activity with respect to ATP
653078
Pseudomonas denitrificans (nomen rejiciendum)
dADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
hydrogenobyrinic acid a,c-diamide + ATP + Co2+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
ITP + hydrogenobyrinic acid a,c-diamide + Co2+
5% of activity with respect to ATP
653078
Pseudomonas denitrificans (nomen rejiciendum)
IDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
653078
Pseudomonas denitrificans (nomen rejiciendum)
?
Subunits (protein specific)
Subunits
Commentary
Organism
trimer
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
Other publictions for EC 6.6.1.2
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
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)
748769
Moore
Towards a cell factory for vi ...
Bacillus megaterium, Bacillus megaterium DSM 319
New Biotechnol.
31
553-561
2014
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2
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6
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2
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1
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1
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2
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2
-
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-
-
-
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-
1
-
-
1
-
-
716791
Romao
Evolution in a family of chela ...
Desulfovibrio vulgaris, Salmonella enterica
Proc. Natl. Acad. Sci. USA
108
97-102
2011
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-
2
2
-
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2
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2
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2
-
-
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1
2
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2
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2
2
2
-
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2
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2
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-
1
2
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-
699853
Lundqvist
The AAA(+) motor complex of su ...
Brucella melitensis
J. Struct. Biol.
167
227-234
2009
-
-
1
-
-
-
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-
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-
1
-
3
-
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1
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1
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1
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-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
15270
Debussche
Biosynthesis of the corrin mac ...
Pseudomonas denitrificans (nomen rejiciendum)
J. Bacteriol.
175
7430-7440
1993
-
-
-
-
-
-
-
-
-
-
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1
-
1
-
-
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1
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1
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1
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-
-
-
653078
Debussche
Assay, purification, and chara ...
Pseudomonas denitrificans (nomen rejiciendum)
J. Bacteriol.
174
7445-7451
1992
-
-
-
-
-
-
10
3
-
1
4
1
-
3
-
-
1
-
-
-
1
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
10
-
3
-
1
4
1
-
-
-
1
-
-
1
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-