BRENDA - Enzyme Database

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
EC Number
Inhibitors
Commentary
Organism
Structure
6.6.1.2
adenylyl (beta,gamma-methylene)-diphosphonate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
adenylyl-imidodiphosphate
50% inhibition at 2 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
ADP
50% inhibition at 8 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
AMP
15% inhibition at 10 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Cu2+
50% inhibition at 0.002 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
diphosphate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Fe2+
50% inhibition at 0.05 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
N-ethylmaleimide
50% inhibition at 0.005 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Ni2+
50% inhibition at 0.003 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Zn2+
50% inhibition at 0.0005 mM
Pseudomonas denitrificans (nomen rejiciendum)
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.6.1.2
0.000085
-
hydrogenobyrinic acid a,c-diamide
pH 8.0, 30░C
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
0.0042
-
Co2+
pH 8.0, 30░C
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
0.22
-
ATP
pH 8.0, 30░C
Pseudomonas denitrificans (nomen rejiciendum)
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
6.6.1.2
Mg2+
ATP binding cation
Pseudomonas denitrificans (nomen rejiciendum)
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.6.1.2
38000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
80000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
140000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
258000
-
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
6.6.1.2
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
6.6.1.2
Pseudomonas denitrificans (nomen rejiciendum)
P29933 AND P29934 AND Q9HZQ3 AND P29929
-
-
Purification (Commentary)
EC Number
Commentary
Organism
6.6.1.2
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]
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
6.6.1.2
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.6.1.2
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)
?
6.6.1.2
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)
?
6.6.1.2
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)
?
6.6.1.2
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)
?
6.6.1.2
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
EC Number
Subunits
Commentary
Organism
6.6.1.2
trimer
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
6.6.1.2
adenylyl (beta,gamma-methylene)-diphosphonate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
adenylyl-imidodiphosphate
50% inhibition at 2 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
ADP
50% inhibition at 8 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
AMP
15% inhibition at 10 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Cu2+
50% inhibition at 0.002 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
diphosphate
50% inhibition at 5 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Fe2+
50% inhibition at 0.05 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
N-ethylmaleimide
50% inhibition at 0.005 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Ni2+
50% inhibition at 0.003 mM
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
Zn2+
50% inhibition at 0.0005 mM
Pseudomonas denitrificans (nomen rejiciendum)
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.6.1.2
0.000085
-
hydrogenobyrinic acid a,c-diamide
pH 8.0, 30░C
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
0.0042
-
Co2+
pH 8.0, 30░C
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
0.22
-
ATP
pH 8.0, 30░C
Pseudomonas denitrificans (nomen rejiciendum)
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
6.6.1.2
Mg2+
ATP binding cation
Pseudomonas denitrificans (nomen rejiciendum)
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
6.6.1.2
38000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
80000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
140000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)
6.6.1.2
258000
-
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
6.6.1.2
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)
EC Number
Commentary
Organism
6.6.1.2
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)
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
6.6.1.2
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.6.1.2
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)
?
6.6.1.2
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)
?
6.6.1.2
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)
?
6.6.1.2
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)
?
6.6.1.2
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)
EC Number
Subunits
Commentary
Organism
6.6.1.2
trimer
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Pseudomonas denitrificans (nomen rejiciendum)