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Literature summary for 6.6.1.2 extracted from

  • Debussche, L.; Couder, M.; Thibaut, D.; Cameron, B.; Crouzet, J.; Blanche, F.
    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 (1992), J. Bacteriol., 174, 7445-7451.
    View publication on PubMedView publication on EuropePMC

Inhibitors

Inhibitors Comment Organism Structure
adenylyl (beta,gamma-methylene)-diphosphonate 50% inhibition at 5 mM Pseudomonas denitrificans (nom. rej.)
adenylyl-imidodiphosphate 50% inhibition at 2 mM Pseudomonas denitrificans (nom. rej.)
ADP 50% inhibition at 8 mM Pseudomonas denitrificans (nom. rej.)
AMP 15% inhibition at 10 mM Pseudomonas denitrificans (nom. rej.)
Cu2+ 50% inhibition at 0.002 mM Pseudomonas denitrificans (nom. rej.)
diphosphate 50% inhibition at 5 mM Pseudomonas denitrificans (nom. rej.)
Fe2+ 50% inhibition at 0.05 mM Pseudomonas denitrificans (nom. rej.)
N-ethylmaleimide 50% inhibition at 0.005 mM Pseudomonas denitrificans (nom. rej.)
Ni2+ 50% inhibition at 0.003 mM Pseudomonas denitrificans (nom. rej.)
Zn2+ 50% inhibition at 0.0005 mM Pseudomonas denitrificans (nom. rej.)

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.000085
-
hydrogenobyrinic acid a,c-diamide pH 8.0, 30°C Pseudomonas denitrificans (nom. rej.)
0.0042
-
Co2+ pH 8.0, 30°C Pseudomonas denitrificans (nom. rej.)
0.22
-
ATP pH 8.0, 30°C Pseudomonas denitrificans (nom. rej.)

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ ATP binding cation Pseudomonas denitrificans (nom. rej.)

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
38000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE Pseudomonas denitrificans (nom. rej.)
80000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE Pseudomonas denitrificans (nom. rej.)
140000
-
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE Pseudomonas denitrificans (nom. rej.)
258000
-
gel filtration Pseudomonas denitrificans (nom. rej.)

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
hydrogenobyrinic acid a,c-diamide + ATP + Co2+ Pseudomonas denitrificans (nom. rej.)
-
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas denitrificans (nom. rej.) P29933 AND P29934 AND Q9HZQ3 AND P29929
-
-

Purification (Commentary)

Purification (Comment) 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 (nom. rej.)

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment 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 (nom. rej.)

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + hydrogenobyrinic acid a,c-diamide + Co2+
-
Pseudomonas denitrificans (nom. rej.) ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
?
CTP + hydrogenobyrinic acid a,c-diamide + Co2+ 45% of activity with respect to ATP Pseudomonas denitrificans (nom. rej.) CDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
?
dATP + hydrogenobyrinic acid a,c-diamide + Co2+ 65% of activity with respect to ATP Pseudomonas denitrificans (nom. rej.) dADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
?
hydrogenobyrinic acid a,c-diamide + ATP + Co2+
-
Pseudomonas denitrificans (nom. rej.) ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
?
ITP + hydrogenobyrinic acid a,c-diamide + Co2+ 5% of activity with respect to ATP Pseudomonas denitrificans (nom. rej.) IDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
-
?

Subunits

Subunits Comment Organism
trimer 1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE Pseudomonas denitrificans (nom. rej.)

Synonyms

Synonyms Comment Organism
cobalt chelatase
-
Pseudomonas denitrificans (nom. rej.)
cobaltochelatase
-
Pseudomonas denitrificans (nom. rej.)
CobN-CobST
-
Pseudomonas denitrificans (nom. rej.)
CobNST
-
Pseudomonas denitrificans (nom. rej.)
gene cobN/gene cobS cobaltochelatase
-
Pseudomonas denitrificans (nom. rej.)
holocobalamin synthase
-
Pseudomonas denitrificans (nom. rej.)
hydrogenobyrinic acid a,c-diamide cobaltochelatase
-
Pseudomonas denitrificans (nom. rej.)