Information on EC 6.3.5.11 - cobyrinate a,c-diamide synthase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
6.3.5.11
-
RECOMMENDED NAME
GeneOntology No.
cobyrinate a,c-diamide synthase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2 ATP + cobyrinate + 2 L-glutamine + 2 H2O = 2 ADP + 2 phosphate + cobyrinate a,c-diamide + 2 L-glutamate
show the reaction diagram
ATP + cobyrinate + L-glutamine + H2O = ADP + phosphate + cobyrinate c-monamide + L-glutamate
show the reaction diagram
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-
-
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ATP + cobyrinate c-monamide + L-glutamine + H2O = ADP + phosphate + cobyrinate a,c-diamide + L-glutamate
show the reaction diagram
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
cob(II)yrinate a,c-diamide biosynthesis I (early cobalt insertion)
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-
Porphyrin and chlorophyll metabolism
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vitamin B12 metabolism
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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co-expression of the cobA gene from Propionibacterium freudenreichii and the cbiA, -C, -D, -E, -T, -F, -G, -H, -J, -K, -L, and -P genes from Salmonella enterica serovar typhimurium in Escherichia coli result in the production of cobyrinic acid a,c-diamide. Strains that have neither cbiP nor cbiA synthesized 1-desmethylcobyrinic acid even in the presence of cbiD, suggesting that CbiA and CbiP are necessary for CbiD activity
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 ATP + (aq)2cobyrinate + 2 L-glutamine + 2 H2O
2 ADP + 2 phosphate + (aq)2cobyrinate a,c-diamide + 2 L-glutamate
show the reaction diagram
-
76% of the initial velocity with cobyrinate
-
-
?
2 ATP + (CN,aq)cobyrinate + 2 L-glutamine + 2 H2O
2 ADP + 2 phosphate + (CN,aq)cobyrinate a,c-diamide + 2 L-glutamate
show the reaction diagram
-
-
-
-
?
2 ATP + cobyrinate + 2 ammonia + 2 H2O
2 ADP + 2 phosphate + cobyrinate a,c-diamide
show the reaction diagram
2 ATP + cobyrinate + 2 L-glutamine + 2 H2O
2 ADP + 2 phosphate + cobyrinate a,c-diamide + 2 L-glutamate
show the reaction diagram
2 ATP + cobyrinate g-monoamide + L-glutamine + 2 H2O
2 ADP + phosphate + cobyrinate c,g-diamide + L-glutamate
show the reaction diagram
-
145% of the initial velocity with cobyrinate
product is 83% cobyrinate c,g-diamide, 17% cobyrinate a,c,g-triamide
-
?
2 ATP + hydrogenobyrinate + 2 L-glutamine + 2 H2O
2 ADP + 2 phosphate + hydrogenobyrinate a,c-diamide + 2 L-glutamate
show the reaction diagram
-
-
-
-
?
ATP + (CN,aq)cobyrinate c-monoamide + L-glutamine + H2O
ADP + phosphate + (CN,aq)cobyrinate a,c-diamide + L-glutamate
show the reaction diagram
-
-
-
-
?
ATP + cobyrinate a-monoamide + L-glutamine + H2O
ADP + phosphate + cobyrinate a,c-diamide + L-glutamate
show the reaction diagram
-
81% of the initial velocity with cobyrinate
product is 100% cobyrinate a,c-diamide
-
?
ATP + cobyrinate c-monoamide + L-glutamine + H2O
ADP + phosphate + cobyrinate a,c-diamide + L-glutamate
show the reaction diagram
-
19% of the initial velocity with cobyrinate
product is 100% cobyrinate a,c-diamide
-
?
ATP + hydrogenobyrinate c-monoamide + L-glutamine + H2O
ADP + phosphate + hydrogenobyrinate a,c-diamide + L-glutamate
show the reaction diagram
-
19% of the initial velocity with cobyrinate
-
-
?
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(aq)2cobyrinate
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substrate inhibition above 0.3 mM
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.25
(aq)2cobyrinate
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or above, pH 7.6, 30C
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0.16
(CN,aq)cobyrinate
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pH 7.6, 30C
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0.071
(CN,aq)cobyrinate c-monoamide
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pH 7.6, 30C
-
12 - 26.2
ammonia
0.00041
Hydrogenobyrinate
-
pH 7.6, 30C
0.00021
hydrogenobyrinate c-monoamide
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pH 7.6, 30C
0.0203 - 0.053
L-glutamine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.18
ammonia
Salmonella enterica subsp. enterica serovar Typhimurium
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pH 7.7, 30C
0.13
ATP
Salmonella enterica subsp. enterica serovar Typhimurium
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pH 7.7, 30C
0.16
cobyrinate
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 7.7, 30C
0.16
L-glutamine
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 7.7, 30C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.9
ammonia
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 7.7, 30C
753
48.2
ATP
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 7.7, 30C
4
216
cobyrinate
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 7.7, 30C
9454
3.04
L-glutamine
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 7.7, 30C
102
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1320
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pH 7.6, 30C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46000
-
2 * 46000, SDS-PAGE
50000
-
1 * 50039, calculated, x * 50000, SDS-PAGE
50039
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1 * 50039, calculated, x * 50000, SDS-PAGE
86000
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gel fitlration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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2 * 46000, SDS-PAGE
monomer
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
from recombinant Pseudomonas denitrificans
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D45N
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below 0.02% of the amidation activity of wild-type, no change in the order of amidation
D48N
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0.2% of the amidation activity of wild-type, no change in the order of amidation
D97N
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32.4% of the amidation activity of wild-type, no change in the order of amidation
E90Q
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below 0.02% of the amidation activity of wild-type, no change in the order of amidation
L47A
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10.6% of the amidation activity of wild-type, specific decrease in the affinity of the enzyme for the c-monoamide intermediate
Y46A
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3.8% of the amidation activity of wild-type, specific decrease in the affinity of the enzyme for the c-monoamide intermediate
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
co-expression of the cobA gene from Propionibacterium freudenreichii and the cbiA, -C, -D, -E, -T, -F, -G, -H, -J, -K, -L, and -P genes from Salmonella enterica serovar typhimurium in Escherichia coli result in the production of cobyrinic acid a,c-diamide. Strains that have neither cbiP nor cbiA synthesized 1-desmethylcobyrinic acid even in the presence of cbiD, suggesting that CbiA and CbiP are necessary for CbiD activity
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