Information on EC 2.5.1.57 - N-acylneuraminate-9-phosphate synthase

Word Map on EC 2.5.1.57
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
2.5.1.57
-
RECOMMENDED NAME
GeneOntology No.
N-acylneuraminate-9-phosphate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
phosphoenolpyruvate + N-acyl-D-mannosamine 6-phosphate + H2O = N-acylneuraminate 9-phosphate + phosphate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
condensation
Phosphorylation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
CMP-N-acetylneuraminate biosynthesis I (eukaryotes)
-
-
Amino sugar and nucleotide sugar metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphoenolpyruvate:N-acyl-D-mannosamine-6-phosphate 1-(2-carboxy-2-oxoethyl)transferase
Acts on N-glycoloyl and N-acetyl-derivatives.
CAS REGISTRY NUMBER
COMMENTARY hide
9031-58-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
neuB gene product amino acid sequence accession. no.
-
-
Manually annotated by BRENDA team
mouse, neuB gene product accession. no.
SwissProt
Manually annotated by BRENDA team
sheep
-
-
Manually annotated by BRENDA team
Sprague-Dawley
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D-mannose 6-phosphate + ?
?
show the reaction diagram
-
-
-
-
?
D-mannose 6-phosphate + phosphoenolpyruvate + H2O
2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid 9-phosphate + phosphate
show the reaction diagram
-
-
-
-
?
D-mannose 6-phosphate + phosphoenolpyruvate + H2O
2-keto-3-deoxy-D-glycero-D-galactononoic acid + phosphate
show the reaction diagram
-
-
?
N-acetyl-D-mannosamine 6-phosphate + phosphoenolpyruvate + H2O
N-acetyl-neuraminate 9-phosphate + phosphate
show the reaction diagram
N-acetyl-D-mannosamine 6-phosphate + phosphoenolpyruvate + H2O
N-acetylneuraminate 9-phosphate + phosphate
show the reaction diagram
-
-
-
-
?
N-acetyl-D-mannosamine 6-phosphate + phosphoenolpyruvate + H2O
N-acetylneuraminic acid 9-phosphate + phosphate
show the reaction diagram
-
-
-
?
N-acyl-D-mannosamine 6-phosphate + phosphoenolpyruvate + H2O
N-acylneuraminate 9-phosphate + phosphate
show the reaction diagram
N-acyl-D-mannosamine 6-phosphate + phosphoenolpyruvate + H2O
N-acylneuraminic acid 9-phosphate + phosphate
show the reaction diagram
N-glycolyl-D-mannosamine 6-phosphate + phosphoenolpyruvate + H2O
N-glycolyl-neuraminate 9-phosphate + phosphate
show the reaction diagram
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
N-acetyl-D-mannosamine 6-phosphate + phosphoenolpyruvate + H2O
N-acetyl-neuraminate 9-phosphate + phosphate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
1 mM, 30% increase of activity
Co2+
-
-
Zn2+
-
-
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5,5'-dithio-bis(2-nitrobenzoic acid)
-
-
Ba2+
-
1 mM, 93% decrease of activity
Ca2+
-
1 mM, 76% decrease of activity
Co2+
-
1 mM, 15% decrease of activity
Cu2+
-
1 mM, 68% decrease of activity
Fe2+
-
1 mM, 79% decrease of activity
Mg2+
-
12.5 mM Mg2+ decrease the ability for N-acetylneuraminate 9-phosphate synthesis by almost half, while the 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid 9-phosphate synthetic activity drops by 20%
N-ethylmaleimide
-
-
Ni2+
-
1 mM, 24% decrease of activity
Zn2+
-
1 mM, 87% decrease of activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5 - 5
D-mannose 6-phosphate
0.035 - 1.72
N-acetyl-D-mannosamine 6-phosphate
1.6
N-Glycolyl-D-mannosamine 6-phosphate
-
-
0.1
phosphoenolpyruvate
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008 - 0.67
D-mannose 6-phosphate
0.013 - 1.31
N-acetyl-D-mannosamine 6-phosphate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0013
-
smooth muscle cells
0.0015
-
endothelial cells
0.2
-
-
0.22
-
in the presence of 12.5 mM MnCl2, 8.3 mM phosphoenolpyruvate and 12.5 mM D-mannose 6-phosphate
0.26
-
-
11.7
-
in the presence of 12.5 mM MnCl2, 8.3 mM phosphoenolpyruvate and 2.5 mM N-acyl-D-mannosamine 6-phosphate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 10
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 70
-
-
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.7
-
SDS-PAGE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
-
SDS-PAGE
42000
-
SDS-PAGE
42360
-
calculated from sequence of cDNA; ESI mass spectrometry
75000
-
gel filtration
80000
gel filtration
82000
-
gel filtration
86000
-
SDS-PAGE
100100
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer or trimer
-
the molecular weight is more than the size of a dimer, but smaller than a trimer
homodimer
-
2 * 41000, gel filtration; 2 * 43000, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 65
-
the enzyme loses 43% of its initial KDN-9-P synthetic activity after 10 min at 37C or 55C, treatment for 10 min at 65C results in retention of 26% and 39% of N-acetylneuraminate 9-phosphate and 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid-9-phosphate activity respectively
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
protected from inactivation by presence of phosphoenolpyruvate
-
purified enzyme is unstable
-
purified enzyme is unstable to freezing and thawing
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-18C, crude extract can be stored
-
-20C, crude extract stable for at least 1 month
-
0C, loses approximately 25% of the activity during a 24-h period
-
4C, crude extract retains 60% of enzyme activity when stored for 1 week
-
4C, purified enzyme loses activity completely after 1 day in phosphate buffer
-
4C, purified enzyme loses approximately half of its activity in 3 days
-
4C, purified enzyme, 50% of enzyme activity remains after 2 days in phosphate buffer containing 5 mM MgCl2, enzyme activity is lost entirely after 3 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HisTrap column chromatography and HiTrap SP column chromatography
-
Ni2+ chelating Sepharose column chromatography
-
Ni2+-Sepharose Fast Flow column chromatography
-
recombinant enzyme
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; cloned and expressed in either bacterial or baculoviral expression systems
cDNA cloned by PCR, transiently expressed in HeLa cells
expressed in Arabidopsis thaliana
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21 cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
M42L
-
mutant shows the additional 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid 9-phosphate synthase activity like the wild type enzyme
M42T
-
the mutation abolishes the 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid 9-phosphate synthase activity completely without compromising the Neu5Ac-9-P synthase activity
additional information
-
by replacing the antifreeze-like domain of the human NeuNAc-9-P synthase with the mouse antifreeze-like domain, the chimeric protein retains almost half of the ability of the human enzyme for 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid-9-phosphate synthesis while the N-acetylneuraminate 9-phosphate production is reduced to less than 10%, the chimeric enzyme is less resistant to high temperatures compared to the wild type enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
synthesis of CMP-N-acetylneuraminic acid in Sf9 expression system by coexpression of enzyme with UDP-GlcNAc-2-epimerase/ManNAc kinase and CMP-sialic acid synthetase plus addition of N-acetylglucosamine
Show AA Sequence (124 entries)
Please use the Sequence Search for a certain query.