Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 6.3.5.11 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 in Escherichia coli Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

Protein Variants Comment Organism
D45N below 0.02% of the amidation activity of wild-type, no change in the order of amidation Salmonella enterica subsp. enterica serovar Typhimurium
D48N 0.2% of the amidation activity of wild-type, no change in the order of amidation Salmonella enterica subsp. enterica serovar Typhimurium
D97N 32.4% of the amidation activity of wild-type, no change in the order of amidation Salmonella enterica subsp. enterica serovar Typhimurium
E90Q below 0.02% of the amidation activity of wild-type, no change in the order of amidation Salmonella enterica subsp. enterica serovar Typhimurium
L47A 10.6% of the amidation activity of wild-type, specific decrease in the affinity of the enzyme for the c-monoamide intermediate Salmonella enterica subsp. enterica serovar Typhimurium
Y46A 3.8% of the amidation activity of wild-type, specific decrease in the affinity of the enzyme for the c-monoamide intermediate Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00074
-
cobyrinate pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
0.0027
-
ATP pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
0.053
-
L-glutamine pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
26.2
-
ammonia pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
1 * 50039, calculated, x * 50000, SDS-PAGE Salmonella enterica subsp. enterica serovar Typhimurium
50039
-
1 * 50039, calculated, x * 50000, SDS-PAGE Salmonella enterica subsp. enterica serovar Typhimurium
53000
-
gel filtration Salmonella enterica subsp. enterica serovar Typhimurium

Organism

Organism UniProt Comment Textmining
Salmonella enterica subsp. enterica serovar Typhimurium
-
LT2
-

Reaction

Reaction Comment Organism Reaction ID
2 ATP + cobyrinate + 2 L-glutamine + 2 H2O = 2 ADP + 2 phosphate + cobyrinate a,c-diamide + 2 L-glutamate 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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 ATP + cobyrinate + 2 ammonia + 2 H2O
-
Salmonella enterica subsp. enterica serovar Typhimurium 2 ADP + 2 phosphate + cobyrinate a,c-diamide
-
?
2 ATP + cobyrinate + 2 L-glutamine + 2 H2O
-
Salmonella enterica subsp. enterica serovar Typhimurium 2 ADP + 2 phosphate + cobyrinate a,c-diamide + 2 L-glutamate the hydrolysis of glutamine and the synthesis of the cobyrinic acid a,c-diamide product are uncoupled. CbiA catalyzes the sequential amidation of the c- and a-carboxylate groups of cobyrinic acid via the formation of a phosphorylated intermediate. 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 ?
additional information CbiA is able to hydrolyze glutamine in the absence of the other two substrates. The addition of ATP alone decreases the Km for glutamine by a factor of 2. Cobyrinic acid enhances the kcat for the hydrolysis of glutamine about 2fold and decreases the Km for glutamine by 20fold. The decrease in the Km for glutamine is about 200fold when both ATP and cobyrinic acid are present in the assay solution. There is no change in the Km for glutamine when ADP is added with cobyrinic acid Salmonella enterica subsp. enterica serovar Typhimurium ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 50039, calculated, x * 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
0.13
-
ATP pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
0.16
-
L-glutamine pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
0.16
-
cobyrinate pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
0.18
-
ammonia pH 7.7, 30°C 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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.04
-
L-glutamine pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
6.9
-
ammonia pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
48.2
-
ATP pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium
216
-
cobyrinate pH 7.7, 30°C Salmonella enterica subsp. enterica serovar Typhimurium