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Literature summary for 6.3.5.10 extracted from

  • Blanche, F.; Couder, M.; Debussche, L.; Thibaut, D.; Cameron, B.; Cruzet, J.
    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 (1991), J. Bacteriol., 173, 6046-6051.
    View publication on PubMedView publication on EuropePMC
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Cloned(Commentary)

Cloned (Comment) Organism
cobQ gene Pseudomonas denitrificans (nom. rej.)

General Stability

General Stability Organism
20% w/v and 1 mM dithiothreitol are necessary to stabilize enzyme during purification Pseudomonas denitrificans (nom. rej.)

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information
-
 Pseudomonas denitrificans (nom. rej.)
0.00104
-
 adenosylcobyrinic acid a,c-diamide pH 7.5 Pseudomonas denitrificans (nom. rej.)
0.00121
-
 adenosylcobyrinic acid tetraamide pH 7.5 Pseudomonas denitrificans (nom. rej.)
0.00122
-
 adenosylcobyrinic acid triamide pH 7.5 Pseudomonas denitrificans (nom. rej.)
0.00125
-
 adenosylcobyrinic acid pentaamide pH 7.5 Pseudomonas denitrificans (nom. rej.)
0.011
-
 ATP pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate Pseudomonas denitrificans (nom. rej.)
0.033
-
 L-glutamine pH 7.5, adenosylcobyrinic acid triamide as substrate Pseudomonas denitrificans (nom. rej.)
0.035
-
 ATP pH 7.5, adenosylcobyrinic acid triamide as substrate Pseudomonas denitrificans (nom. rej.)
0.045
-
 L-glutamine pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate Pseudomonas denitrificans (nom. rej.)
0.049
-
 L-glutamine pH 7.5, adenosylcobyrinic acid pentaamide as substrate Pseudomonas denitrificans (nom. rej.)
0.064
-
 L-glutamine pH 7.5, adenosylcobyrinic acid tetraamide as substrate Pseudomonas denitrificans (nom. rej.)
0.088
-
 ATP pH 7.5, adenosylcobyrinic acid pentaamide as substrate Pseudomonas denitrificans (nom. rej.)
0.12
-
 ATP pH 7.5, adenosylcobyrinic acid tetraamide as substrate Pseudomonas denitrificans (nom. rej.)
20.4
-
 ammonia pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate Pseudomonas denitrificans (nom. rej.)

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ Mg2+-dependent Pseudomonas denitrificans (nom. rej.)

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52000
-
 predicted from DNA sequence of the cobQ gene Pseudomonas denitrificans (nom. rej.)
57000
-
 2 * 57000, SDS-PAGE Pseudomonas denitrificans (nom. rej.)
97300
-
 gel filtration Pseudomonas denitrificans (nom. rej.)

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O Pseudomonas denitrificans (nom. rej.) biosynthesis of vitamin B12, cobalamin biosynthetic pathway ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O Pseudomonas denitrificans (nom. rej.) SC510 RifT biosynthesis of vitamin B12, cobalamin biosynthetic pathway ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas denitrificans (nom. rej.)
-
-
-
Pseudomonas denitrificans (nom. rej.)
-
 recombinant strain SC510 Rifr(pXL617)
-

Oxidation Stability

Oxidation Stability Organism
sensitive to oxidation Pseudomonas denitrificans (nom. rej.)

Purification (Commentary)

Purification (Comment) Organism
45fold Pseudomonas denitrificans (nom. rej.)

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.083
-
 pH 7.5, adenosylcobyrinic acid a,c-diamide as substrate Pseudomonas denitrificans (nom. rej.)

Storage Stability

Storage Stability Organism
-20°C or 0°C, purified enzyme, 0.1 M Tris hydrochloride, pH 7.5, 48 h, 100% loss of activity Pseudomonas denitrificans (nom. rej.)
-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 (nom. rej.)

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) adenosylcobyric acid + ADP + phosphate + L-glutamate
-
 ?
adenosylcobyrinic acid a,c-diamide + ATP + L-glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) SC510 RifT adenosylcobyric acid + ADP + phosphate + L-glutamate
-
 ?
ATP + adenosylcobyrinic acid a,c-diamide + ammonia + H2O ammonia can replace L-glutamine as amide group donor, but L-glutamine is preferred Pseudomonas denitrificans (nom. rej.) ADP + phosphate + adenosylcobyric acid + ?
-
 ?
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O biosynthesis of vitamin B12, cobalamin biosynthetic pathway Pseudomonas denitrificans (nom. rej.) ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
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 Pseudomonas denitrificans (nom. rej.) ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
ATP + adenosylcobyrinic acid a,c-diamide + L-glutamine + H2O biosynthesis of vitamin B12, cobalamin biosynthetic pathway Pseudomonas denitrificans (nom. rej.) SC510 RifT ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
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 Pseudomonas denitrificans (nom. rej.) SC510 RifT ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
ATP + adenosylcobyrinic acid pentaamide + L-glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
ATP + adenosylcobyrinic acid tetraamide + L-glutamine + H2O two-step amidation Pseudomonas denitrificans (nom. rej.) ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
ATP + adenosylcobyrinic acid triamide + L-glutamine + H2O three-step amidation Pseudomonas denitrificans (nom. rej.) ADP + phosphate + adenosylcobyric acid + L-glutamate
-
 ?
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) cobinamide + ADP + phosphate + L-glutamate
-
 ?
cobinic acid a,c-diamide + ATP + L-glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) SC510 RifT cobinamide + ADP + phosphate + L-glutamate
-
 ?
additional information no amidation of adenosylcobyric acid, hydrogenobyrinic acid a,c-diamide, monocyano or diaqua form of cobyrinic acid a,c-diamide Pseudomonas denitrificans (nom. rej.) ?
-
 ?
additional information no amidation of adenosylcobyric acid, hydrogenobyrinic acid a,c-diamide, monocyano or diaqua form of cobyrinic acid a,c-diamide Pseudomonas denitrificans (nom. rej.) SC510 RifT ?
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 57000, SDS-PAGE Pseudomonas denitrificans (nom. rej.)
homodimer 2 * 57000, SDS-PAGE Pseudomonas denitrificans (nom. rej.)

Synonyms

Synonyms Comment Organism
CobQ
-
 Pseudomonas denitrificans (nom. rej.)

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 broad pH-optimum around Pseudomonas denitrificans (nom. rej.)

Cofactor

Cofactor Comment Organism Structure
ATP ATP-dependent Pseudomonas denitrificans (nom. rej.)

pI Value

Organism Comment pI Value Maximum pI Value
Pseudomonas denitrificans (nom. rej.) isoelectric focusing
-
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