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Literature summary for 6.3.5.9 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
-
 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 Comment 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 Comment Organism Structure
Mg2+ amidation reaction is Mg2+ dependent Pseudomonas denitrificans (nom. rej.)

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas denitrificans (nom. rej.)
-
-
-
Pseudomonas denitrificans (nom. rej.) SC510 RifT
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas denitrificans (nom. rej.)

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O
-
 Pseudomonas denitrificans (nom. rej.) 5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
 ?
5'-deoxy-5'-adenosyl-cobyrinic acid + 2 ATP + 2 glutamine + 2 H2O cobalamin biosynthetic pathway Pseudomonas denitrificans (nom. rej.) 5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + 2 ADP + 2 phosphate + 2 glutamate
-
 ?
5'-deoxy-5'-adenosyl-cobyrinic acid + ATP + ammonia + H2O
-
 Pseudomonas denitrificans (nom. rej.) 5'-deoxy-5'-adenosyl-cobyrinic acid ac-diamide + ADP + phosphate + ?
-
 ?
adenosyl-cobyrinic acid pentaamide + ATP + glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) ADP + glutamate + phosphate + ?
-
 ?
adenosyl-cobyrinic acid tetraamide + ATP + glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) ADP + glutamate + phosphate + ?
-
 ?
adenosyl-cobyrinic acid triamide + ATP + glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) ADP + glutamate + phosphate + ?
-
 ?
ATP + adenosyl-cobyrinic acid a,c-diamide + L-glutamine + H2O
-
 Pseudomonas denitrificans (nom. rej.) ADP + phosphate + ?
-
 ?
additional information adenosyl-cobyric acid and hydrogenobyrinic acid a,c-diamide are no substrates Pseudomonas denitrificans (nom. rej.) ?
-
 ?

Subunits

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

Synonyms

Synonyms Comment Organism
cobyric acid synthase
-
 Pseudomonas denitrificans (nom. rej.)

pH Optimum

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

pI Value

Organism Comment pI Value Maximum pI Value
Pseudomonas denitrificans (nom. rej.)
-
 5.9 5.6