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Literature summary for 6.3.5.9 extracted from

  • Fresquet, V.; Williams, L.; Raushel, F.M.
    Mechanism of cobyrinic acid a,c-diamide synthetase from Salmonella typhimurium LT2 (2004), Biochemistry, 43, 10619-10627.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

Protein Variants Comment Organism
D45N less than 0.2% of amidation activity with glutamine and with NH4+, compared to wild-type values Salmonella enterica subsp. enterica serovar Typhimurium
D48N less than 0.3% of amidation activity with glutamine and with NH4+, compared to wild-type values Salmonella enterica subsp. enterica serovar Typhimurium
D97N 32.4% of wild-type amidation activity with glutamine, 16.5% of wild-type amidation activity with NH4+ Salmonella enterica subsp. enterica serovar Typhimurium
E90Q less than 0.02% of amidation activity with glutamine and with NH4+, compared to wild-type values Salmonella enterica subsp. enterica serovar Typhimurium
L47A 10.6% of wild-type amidation activity with glutamine, 44% of wild-type amidation activity with NH4+, mutation specifically decreases the affinity of the enzyme to the cobyrinic acid c-monoamide Salmonella enterica subsp. enterica serovar Typhimurium
Y46A 3.8% of wild-type amidation activity with glutamine, 2% of wild-type amidation activity with NH4+, mutation specifically decreases the affinity of the enzyme to the cobyrinic acid c-monoamide Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0003
-
cobyrinic acid c-monoamide mutant enzyme D97N Salmonella enterica subsp. enterica serovar Typhimurium
0.00041
-
cobyrinic acid c-monoamide wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.00074
-
cobyrinic acid pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.0027
-
cobyrinic acid c-monoamide mutant enzyme L47A Salmonella enterica subsp. enterica serovar Typhimurium
0.0027
-
ATP pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.011
-
cobyrinic acid c-monoamide mutant enzyme Y46A Salmonella enterica subsp. enterica serovar Typhimurium
0.053
-
glutamine pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
26.2
-
NH4+ pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
1 * 50000, SDS-PAGE Salmonella enterica subsp. enterica serovar Typhimurium
55000
-
gel filtration Salmonella enterica subsp. enterica serovar Typhimurium

Organism

Organism UniProt Comment Textmining
Salmonella enterica subsp. enterica serovar Typhimurium
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cobyrinic acid + ATP + glutamine model for catalytic mechanism in which CbiA catalyzes the amidation of the c-carboxylate and then the intermediate is released into solution and binds to the same catalytic site for the amidation of the a-carboxylate Salmonella enterica subsp. enterica serovar Typhimurium cobyrinic acid a,c-diamide + ADP + phosphate + glutamate
-
?
cobyrinic acid + ATP + NH4+ model for catalytic mechanism in which CbiA catalyzes the amidation of the c-carboxylate and then the intermediate is released into solution and binds to the same catalytic site for the amidation of the a-carboxylate Salmonella enterica subsp. enterica serovar Typhimurium cobyrinic acid a,c-diamide + ADP + phosphate
-
?
cobyrinic acid c-monoamide + ATP + glutamine
-
Salmonella enterica subsp. enterica serovar Typhimurium cobyrinic acid a,c-diamide + ADP + phosphate + glutamate
-
?
cobyrinic acid c-monoamide + ATP + NH4+
-
Salmonella enterica subsp. enterica serovar Typhimurium cobyrinic acid a,c-diamide + ADP + phosphate
-
?

Subunits

Subunits Comment Organism
monomer 1 * 50000, SDS-PAGE Salmonella enterica subsp. enterica serovar Typhimurium

Synonyms

Synonyms Comment Organism
CbiA
-
Salmonella enterica subsp. enterica serovar Typhimurium

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.008
-
cobyrinic acid c-monoamide mutant enzyme Y46A Salmonella enterica subsp. enterica serovar Typhimurium
0.04
-
cobyrinic acid c-monoamide mutant enzyme D97N Salmonella enterica subsp. enterica serovar Typhimurium
0.057
-
cobyrinic acid c-monoamide mutant enzyme L47A Salmonella enterica subsp. enterica serovar Typhimurium
0.13
-
ATP pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.16
-
glutamine pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.16
-
cobyrinic acid pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.18
-
NH4+ pH 7.7, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.2
-
cobyrinic acid c-monoamide wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.8 8
-
Salmonella enterica subsp. enterica serovar Typhimurium