Information on EC 4.1.3.3 - N-acetylneuraminate lyase

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

EC NUMBER
COMMENTARY hide
4.1.3.3
-
RECOMMENDED NAME
GeneOntology No.
N-acetylneuraminate lyase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
N-acetylneuraminate = N-acetyl-D-mannosamine + pyruvate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aldol addition
-
reversible aldol cleavage
aldol condensation
-
-
C-C bond cleavage
-
-
C-C-bond cleavage
condensation
-
-
-
-
elimination
-
-
of an oxo-acid, C-C bond cleavage
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Amino sugar and nucleotide sugar metabolism
-
-
N-acetylneuraminate and N-acetylmannosamine degradation I
-
-
N-acetylneuraminate and N-acetylmannosamine degradation II
-
-
metabolism of amino sugars and derivatives
-
-
SYSTEMATIC NAME
IUBMB Comments
N-acetylneuraminate pyruvate-lyase (N-acetyl-D-mannosamine-forming)
Also acts on N-glycoloylneuraminate, and on O-acetylated sialic acids, other than 4-O-acetylated derivatives.
CAS REGISTRY NUMBER
COMMENTARY hide
9027-60-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
Corynebacterium belfanti
-
-
-
Manually annotated by BRENDA team
strain K1
-
-
Manually annotated by BRENDA team
strain TG1
-
-
Manually annotated by BRENDA team
strain W3110
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
subspecies aureus, methicillin-resistant, gene nanA
UniProt
Manually annotated by BRENDA team
gene nanA
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
-
N-acetylneuraminic acid lyase is a class I aldolase
physiological function
additional information
-
reaction mechanism by crystal structure analysis, and quantum mechanical/molecular mechanical modeling, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
?
show the reaction diagram
-
-
-
-
?
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
pyruvate + (2R,3S)-2,3-dihydroxy-4-oxo-N,N-dipropylbutanamide
show the reaction diagram
-
-
-
-
r
2-deoxy-2-[[(alpha-D-galactopyranosyloxy)acetyl]amino]-D-mannopyranose + pyruvate
3,5-dideoxy-5-[[(alpha-D-galactopyranosyloxy)acetyl]amino]-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
36% yield
-
?
2-deoxy-2-[[(beta-D-galactopyranosyloxy)acetyl]amino]-D-mannopyranose + pyruvate
3,5-dideoxy-5-[[(beta-D-galactopyranosyloxy)acetyl]amino]-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
34% yield
-
?
3-bromo-3-deoxy-D-mannose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 5% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 5% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 10% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 10% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 10% of the activity with N-acetyl-D-mannosamine
-
-
?
3-deoxy-manno-oct-ulosonic acid
arabinose + pyruvate
show the reaction diagram
-
-
-
-
?
4-O-alpha-D-galactopyranosyl-D-mannopyranose + pyruvate
3-deoxy-7-O-alpha-D-galactopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
5% yield
-
?
4-O-beta-D-galactopyranosyl-D-mannopyranose + pyruvate
3-deoxy-7-O-beta-D-galactopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
38% yield
-
?
4-O-beta-D-glucopyranosyl-D-mannopyranose + pyruvate
3-deoxy-7-O-beta-D-glucopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
35% yield
-
?
5-N-acetyl-4-O-acetylneuraminic acid
?
show the reaction diagram
-
22% the activity with N-acetylneuraminic acid
-
?
5-N-acetyl-9-O-acetylneuraminic acid
?
show the reaction diagram
5-O-alpha-D-galactopyranosyl-alpha-D-mannofuranose + pyruvate
3-deoxy-8-O-alpha-D-galactopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
62% yield
-
?
5-O-beta-D-galactopyranosyl-alpha-D-mannofuranose + pyruvate
3-deoxy-8-O-beta-D-galactopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
85% yield
-
?
5-O-methyl-N-acetyl-D-mannosamine + pyruvate
8-O-methyl-N-acetylneuraminate
show the reaction diagram
-
-
-
-
6-O-alpha-D-galactopyranosyl-D-mannopyranose + pyruvate
3-deoxy-9-O-alpha-D-galactopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
81% yield
-
?
6-O-alpha-D-glucopyranosyl-D-mannopyranose + pyruvate
3-deoxy-9-O-alpha-D-glucopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
65% yield
-
?
6-O-alpha-D-mannopyranosyl-D-mannopyranose + pyruvate
3-deoxy-9-O-alpha-D-mannopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
78% yield
-
?
6-O-beta-D-glucopyranosyl-D-mannopyranose + pyruvate
3-deoxy-9-O-beta-D-glucopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
show the reaction diagram
-
-
83% yield
-
?
D-3-deoxy-manno-2-octulosonic acid
D-arabinose + pyruvate
show the reaction diagram
-
-
-
-
r
D-arabinose + pyruvate
D-3-deoxy-manno-2-octulosonic acid
show the reaction diagram
-
wild-type enzyme: 20% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 40% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 65% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 40% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 85% of the activity with N-acetyl-D-mannosamine
-
-
r
D-glucose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 40% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 40% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 20% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 25% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 20% of the activity with N-acetyl-D-mannosamine
-
-
?
D-gulose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 10% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 110% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 90% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 70% of the activity with N-acetyl-D-mannosamine
-
-
?
D-mannosamine + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 80% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 90% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 165% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 190% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 170% of the activity with N-acetyl-D-mannosamine
-
-
?
D-mannose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 30% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 15% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 70% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 80% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 75% of the activity with N-acetyl-D-mannosamine
-
-
?
D-mannose + pyruvic acid
2-keto-3-deoxy-D-glycero-D-galacto-nonopyranulosonic acid
show the reaction diagram
-
-
-
-
?
D-talose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 60% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 50% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 105% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 80% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 70% of the activity with N-acetyl-D-mannosamine
-
-
?
D-xylose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 45% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 40% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 55% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 25% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 25% of the activity with N-acetyl-D-mannosamine
-
-
?
L-3-deoxy-manno-2-octulosonic acid
L-arabinose + pyruvate
show the reaction diagram
-
-
-
-
r
L-arabinose + ?
3-deoxy-L-manno-oct-2-ulosonic acid
show the reaction diagram
-
-
?
L-arabinose + pyruvate
L-3-deoxy-manno-2-octulosonic acid
show the reaction diagram
-
wild-type enzyme: 10% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 35% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 70% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 40% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 80% of the activity with N-acetyl-D-mannosamine
-
-
r
L-gulose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 70% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 35% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 15% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 15% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 15% of the activity with N-acetyl-D-mannosamine
-
-
?
L-mannose + pyruvate
?
show the reaction diagram
-
wild-type enzyme: 80% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 110% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 95% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 95% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 95% of the activity with N-acetyl-D-mannosamine
-
-
?
Methoxyneuraminic acid
?
show the reaction diagram
-
-
-
-
?
N,O-Diacetylneuramininic acid
?
show the reaction diagram
-
-
-
-
?
N-Acetyl-4-O-acetylneuraminic acid
?
show the reaction diagram
-
-
-
-
?
N-Acetyl-7-O-acetylneuraminic acid
?
show the reaction diagram
-
-
-
-
?
N-Acetyl-8-O-acetylneuraminic acid
?
show the reaction diagram
-
-
-
-
?
N-Acetyl-9-O-acetylneuraminic acid
?
show the reaction diagram
-
-
-
-
?
N-acetyl-D-mannosamine + pyruvate
N-acetyl-D-neuraminic acid
show the reaction diagram
-
-
-
-
r
N-acetyl-D-mannosamine + pyruvate
N-acetylneuraminate
show the reaction diagram
N-Acetyl-D-mannosamine + pyruvate
N-Acetylneuraminic acid
show the reaction diagram
N-acetyl-D-mannosamine + pyruvic acid
N-acetyl-D-neuraminic acid
show the reaction diagram
-
-
-
-
-
N-acetyl-D-neuraminic acid
N-acetyl-D-mannosamine + pyruvate
show the reaction diagram
N-acetyl-D-neuraminic acid
pyruvate + N-acetyl-D-mannosamine
show the reaction diagram
N-acetyl-L-mannosamine + pyruvate
N-acetyl-L-neuraminic acid
show the reaction diagram
-
wild-type enzyme: 0.2% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I: 0.5% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/V251I/V265I: 1% of the activity with N-acetyl-D-mannosamine, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C: 2% of the activity with N-acetyl-D-mannosamine, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y: 2% of the activity with N-acetyl-D-mannosamine
-
-
r
N-acetyl-L-neuraminic acid
N-acetyl-L-mannosamine + pyruvate
show the reaction diagram
-
-
-
-
r
N-acetyl-neuraminic acid
N-acetyl-D-mannosamine + pyruvate
show the reaction diagram
N-Acetylmannosamine + pyruvate
?
show the reaction diagram
N-acetylmannosamine + pyruvate
N-acetylneuraminic acid
show the reaction diagram
-
-
-
-
r
N-acetylneuraminate
N-acetyl-D-mannosamine + pyruvate
show the reaction diagram
N-Acetylneuraminic acid
?
show the reaction diagram
N-Acetylneuraminic acid
N-Acetyl-D-mannosamine + pyruvate
show the reaction diagram
N-acetylneuraminic acid
N-acetylmannosamine + pyruvate
show the reaction diagram
-
-
-
-
r
N-acetylneuraminic acid
pyruvate + N-acetyl-D-mannosamine
show the reaction diagram
-
-
-
-
r
N-Formylneuraminic acid
?
show the reaction diagram
N-Glycolylneuraminic acid
?
show the reaction diagram
N-Monochloracetylneuraminic acid
?
show the reaction diagram
N-Monofluoracetylneuraminic acid
?
show the reaction diagram
-
-
-
-
?
N-Succinylneuraminic acid
?
show the reaction diagram
-
-
-
-
?
pyruvate + alpha-D-Galp-(1,6)-D-Manp
alpha-D-Galp-(1,9)-D-deoxyneuraminic acid
show the reaction diagram
-
-
-
-
?
additional information
?
-
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 + pyruvate
N-acetylneuraminate
show the reaction diagram
N-acetyl-D-neuraminic acid
N-acetyl-D-mannosamine + pyruvate
show the reaction diagram
N-Acetylmannosamine + pyruvate
?
show the reaction diagram
N-acetylneuraminate
N-acetyl-D-mannosamine + pyruvate
show the reaction diagram
N-Acetylneuraminic acid
?
show the reaction diagram
N-Acetylneuraminic acid
N-Acetyl-D-mannosamine + pyruvate
show the reaction diagram
-
regulation of sialic acid metabolism
-
r
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2R,3R)-2,3,4-trihydroxy-N,N-dipropylbutanamide
-
competitive inhibitor
3-Fluorosialic acid
-
-
4-chloromercuribenzoate
5,5-dithiobis(2-nitrobenzoic acid)
-
50% residual activity at 0.3 mg/ml
5-Diazonium-1-H-tetrazole
Bromopyruvate
Chloropyruvate
-
-
CN-
-
50% residual activity at 20 mM
cyanide
diethyldicarbonate
Diethylpyrocarbonate
-
-
dihydroxyacetone
-
2% residual activity at 10 mM
diisopropylfluorophosphate
DL-glyceraldehyde
-
0% residual activity at 10 mM
Fe3+
-
50% inhibition at 1 mM
Heavy metal ions
-
-
-
hydrogen peroxide
-
0% residual activity at 10 mM, 16% residual activity at 1 mM
Iodine
-
0% residual activity at 0.01 mM, 45% residual activity at 0.001 mM
L-lysine
-
50% inhibition at 0.2 mM, inhibits wild-type enzyme, but not mutant enzymes
Mn2+
-
20% residual activity at 10 mM
N-Acetyl-4-oxo-D-neuraminic acid
N-bromosuccinimide
N-ethylmaleimide
N-Monofluoroacetylneuraminic acid
-
-
NaBH4
o-phenanthroline
-
40% residual activity at 0.1 mM
p-chloromercuribenzoate
phenylmethylsulfonyl fluoride
-
-
pyruvate
Rose bengal
Zn2+
-
1% residual activity at 10 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.39 - 11
(2R,3R,4R)-6-[(aminooxy)carbonyl]-3,4,6-trihydroxy-N-propyltetrahydro-2H-pyran-2-carboxamide
0.35 - 10.6
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
0.2 - 11
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
3.5 - 14.5
D-3-deoxy-manno-2-octulosonic acid
2.4 - 14.1
L-3-deoxy-manno-2-octulosonic acid
12.5
N-acetyl-4-O-acetylneuraminic acid
-
-
4.5
N-Acetyl-7-O-acetylneuraminic acid
-
-
2
N-Acetyl-8-O-acetylneuraminic acid
-
-
7.7 - 245
N-acetyl-D-mannosamine
2.1 - 38
N-acetyl-D-neuraminic acid
3 - 369.4
N-acetyl-L-neuraminic acid
2.8
N-acetyl-neuraminic acid
-
in 0.1 M phosphate buffer, pH 7
0.013 - 6.1
N-acetylneuraminate
0.4 - 1100
N-acetylneuraminic acid
1.75
N-Formylneuraminic acid
-
-
2.2 - 4.6
N-glycolylneuraminic acid
2.7
N-Monochloracetylneuraminic acid
-
-
10
N-Monofluoracetylneuraminic acid
-
-
1.93
N-Succinylneuraminic acid
-
-
1 - 35.1
pyruvate
4.4 - 38
sialic acid
additional information
additional information
-
with substrate N-acetylneuraminic acid increasing Km with increasing pH
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.057 - 10.83
(2R,3R,4R)-6-[(aminooxy)carbonyl]-3,4,6-trihydroxy-N-propyltetrahydro-2H-pyran-2-carboxamide
0.44 - 2.18
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
0.1 - 2.83
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
1.2 - 8.9
D-3-deoxy-manno-2-octulosonic acid
1.1 - 10.4
L-3-deoxy-manno-2-octulosonic acid
4.5 - 11
N-acetyl-D-mannosamine
1.5 - 10.5
N-acetyl-D-neuraminic acid
0.0002 - 0.02
N-acetyl-L-neuraminic acid
0.013 - 16
N-acetylneuraminate
0.42 - 16.7
N-acetylneuraminic acid
1.8 - 2.9
pyruvate
2.833 - 6
sialic acid
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08 - 5.67
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
3579
0.0074 - 0.08
N-acetyl-D-mannosamine
1064
0.0033 - 2
N-acetylneuraminate
730
0.06 - 5.6
N-acetylneuraminic acid
899
0.08 - 0.21
pyruvate
31
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.3
(2R,3R)-2,3,4-trihydroxy-N,N-dipropylbutanamide
-
E192N mutant protein , pH 7.5, 25°C
1
Cu2+
-
-
1
Fe3+
-
-
1.2
pyruvate
-
pH 7.4, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.015
-
N-acetyl-D-mannosamine + pyruvate, 37°C
0.0693
immobilized recombinant enzyme, the activity yield of immobilized enzyme is approximately 10% of the free enzyme
0.14
-
crude cell extract, at 37°C
0.42 - 1.5
-
-
1.2 - 2.2
-
N-acetylneuraminic acid, 25°C
1.5
-
-
2.45
-
crude extract, in 50 mM potassium phosphate buffer, pH 7.5, at 37°C
4.33
purified recombinant wild-type enzyme, pH 7.4, 30°C
6
purified recombinant refolded wild-type enzyme, pH 7.4, 30°C
7.65
-
pH 7.0, 37°C
11
purified recombinant enzyme, pH 8.0, temperature not specified in the publication
12.7
purified recombinant His-taggged enzyme, pH 7.0, 37°C
14.6
-
after purification, at 37°C
35.5
-
N-acetylneuraminic acid
38.7
-
N-acetylneuraminic acid
56.8
-
N-acetylneuraminic acid
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7
6.8
mutant K165-gamma-thialysine
7 - 7.3
-
for both directions
7.2 - 8
-
-
7.2
-
-
7.5 - 8
7.6 - 8
-
-
7.7
-
elimination and condensation
7.8 - 8
-
elimination and condensation
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
pH-activity profiles of the wild-type enzyme, overview
7 - 11.5
-
60% activity at pH 10.0, 97% activity at pH 7.0
7.2 - 8.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
syntesis reaction
70 - 80
-
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 75
-
-
20 - 80
synthesis and hydrolysis reactions, profiles, overview
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
peripheral blood
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aliivibrio salmonicida (strain LFI1238)
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Mycoplasma synoviae (strain 53)
Mycoplasma synoviae (strain 53)
Pasteurella multocida (strain Pm70)
Pasteurella multocida (strain Pm70)
Pasteurella multocida (strain Pm70)
Pasteurella multocida (strain Pm70)
Pasteurella multocida (strain Pm70)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain USA300)
Staphylococcus aureus (strain USA300)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000
-
SDS-PAGE
33992
x * 33992, recombinant His6-tagged wild-type enzyme, mass spectrometry
34600
-
SDS-PAGE
35200
-
-
37317
-
4 * 37317, HPLC/ESI/ion trap
48000
-
2 * 48000, SDS-PAGE
58890
-
GST-tagged enzyme, calculated from amino acid sequence
60000
-
GST-tagged enzyme, SDS-PAGE
61000
-
SDS-PAGE
92000
-
gel filtration
97000
-
gel filtration
98000
-
gel filtration
100000
105000
-
gel filtration
108000
-
gel-filtration
132000
-
gel filtration
135000
-
gel filtration
250000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotetramer
tetramer
trimer
additional information
-
enzyme is a monomer or a tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
L142R mutant, complexed with beta-hydroxypyruvate, hanging drop vapor diffusion method
-
mutant enzyme E192N, in complex with pyruvate, sitting drop vapor diffusion method, using 100 mM Tris-HCl pH 8.2, 200 mM ammonium acetate, 18% PEG 3350
-
sitting drop diffusion method, PEG 3350
-
complexed with sialic acid alditol, 4-deoxy-sialic acid or 4-oxo-sialic acid, microbatch crystallization, hanging drop vapor diffusion method
-
recombinant enzyme in complex with N-acetylneuraminate and acetyl-D-mannosamine/pyruvate, soaking in the mother liquor containing 15% w/v PEG400, then 20% w/v PEG400, and subsequently for 5 min in 25% w/v PEG400 containing 75 mM Neu5Ac, X-ray diffraction structure determination and analysis
-
purified recombinant His-tagged wild-type enzyme, wild-type enzyme with Schiff base and in complex with pyruvate, mutant K164A enzyme, and mutant K164A in complex with N-acetylneuraminic acid and N-glycolylneuraminic acid, hanging drop vapour diffusion method, crystals of wild-type PmNAL from 21% PEG 1000, 150 mM NaCl, and 100 mM Na2HPO4/KH2PO4, pH 6.2. PmNAL K164A mutant in ligand-free from from 30% PEG 200, 100 mM NaCl, and acetate, pH 4.5. Crystals of the PmNAL K164A mutant bound to ligands are grown in 38% PEG 300, 0.01 M CaCl2, and 0.1 M sodium cacodylate, pH 6.5. The sialic acid concentration was 5 mM, X-ray diffraction structure determination and analysis at 1.75-2.10 A resolution, molecular replacement modeling
K0Y2E6
purified recombinant enzyme, hanging drop vapour diffusion method, 0.002-0.0025 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, is mixed with 0.0017-0.0022 ml of reservoir solution containing 25% w/v PEG 3350, 200 mM ammonium sulfate, 100 mM Bis-Tris pH 5.5, equilibration against 1 ml of reservoir solution, 8-20°C, method optimization, X-ray diffraction structure determination and analysis at 1.7 A resolution, molecular replacement
purified recombinant wild-type enzyme and mutants K165C variant and K165-gamma-thialysine, alone or in complex with pyruvate, hanging drop vapour diffusion method, mixing of 0.002 ml of protein solution containing 8 mg/ml protein in 50 mM, pH 7.4, with 0.002 ml of reservoir solution containing 100 mM Tris-HCl, pH 7.0-8.5, 200 mM NaCl, and 18-28% w/v PEG 3350, pyruvate complexes of the wild-type and K165-gamma-thialysine mutant enzymes crystals are soaked in the mother liquor containing 100 mM sodium pyruvate and 15% v/v PEG 400 for 1 min before being sequentially transferred to mother liquor with 5% increments in PEG 400 concentration. The final soak contains the mother liquor containing 100 mM sodium pyruvate and 25% v/v PEG 400, 18°C, X-ray diffraction structure determination and analysis at about 2.0 A resolution, molecular replacement
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
-
purified recombinant enzyme, 24 h, 4°C, different buffers, over 70% activity remains
727332
6 - 7
-
at 75°C stable for more than 15 min
33332
6 - 9
-
at 37°C stable for 24 h
33314
6.5 - 11
-
about 80% activity (synthesis of N-acetylneuraminic acid) after 15 days at pH 8.0-10.0 and 37°C, 60% activity (synthesis of N-acetylneuraminic acid) after 15 days at pH 11.0 and 37°C
713843
7 - 10.5
-
spore-displayed N-acetyl-D-neuraminic acid aldolase is stable for 24 hours at 50°C
713852
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 50
purified recombinant His-tagged enzyme, pH 7.0, completely stable over 140-160 min
45
-
purified recombinant enzyme, 2 h, stable
76
-
in presence of 10 mM substrate stable for 1 min
84
the melting temperature of the wild type enzyme is at 84°C
100
-
complete inactivation after 2 min
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
5-10% activity in the synthetic direction above pH 11.0
-
Amberzyme oxirane resin-immobilized NAL can be used up to five reaction cycles without loss of activity or significant decrease of the conversion rate, the activity yield of immobilized NAL is approximately 10% of the free enzyme
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
sensitive to illumination in presence of Rose Bengal
sensitive to N-bromosuccinimide, H2O2 and iodine
-
33314
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 50 mM Tris/HCl, pH 8, 25 mM MgCl2, 1 mM PMSF, 20% v/v glycerol, stable under prolonged storage
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
100-kDa ultrafiltration step, 65°C heat shock step and Ni2+ chelating affinity chromatography
-
Bio-Gel P-2 gel filtration
-
by FP-based affinity chromatography
expressed in Serratia liquefaciens
-
FPIDA-Co affinity support column chromatography
-
from overproducing strain
-
glutathione-Uniflow Resin column chromatography
-
HisTrap column chromatography
immobilized metal ion affinity chromatography (Ni2+), gel filtration
-
Ni-NTA column chromatography
-
recombinant enzyme from Escherichia coli strain BL21(DE3)
-
recombinant enzyme from Escherichia coli strain BL21(DE3) by anion exchange and hydrophobic interaction chromatography, and gel filtration to over 95% purity
recombinant His-tagged enzyme 7.5fold from Escherichia coli strain Rosetta (DE3) pLys by ultrafiltration, nickel affinity chromatography, and gel filtration, to homogeneity
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
K0Y2E6
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
using a HisTrap_FF column
wild-type and mutant enzymes
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 recombinant Escherichia coli strains capable of expressing N-acetyl-D-glucosamine 2-epimerase and N-acetyl-D-neuraminic acid aldolase are constructed based on a highly efficient temperature-responsive expression system which is safe compared to chemical-induced systems and coupled in N-acetyl-D-neuraminic acid production
-
a Bacillus subtilis outer spore coat protein fusion expressed in Bacillus subtilis WB600 results in a spore-displayed N-acetyl-D-neuraminic acid aldolase
-
cloning and recombinant expression in Escherichia coli strain BL21(DE3)
-
expressed in Bright Yellow 2 (BY2) tobacco cells and Medicago sativa cytosol
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21 (DE3) cells
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli strain BL 21
-
expression in Escherichia coli
gene nal, co-expression in Escherichia coli strain Rosetta (DE3) pLysS with N-acetylglucosamine 2-epimerase from Synechocystis sp. PCC 6803 (snAGE) or Anabaena sp. CH1 (anAGE)
-
gene nanA, DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain BL21(DE3)
gene nanA, recombinant expression of His6-tagged wild-type and mutant enzymes from plasmid pKK223-3 in Escherichia coli strain BL21(DE3)
gene npl, realtime RT-PCR enzyme expression analysis
-
gene ScNAL, sequence comparison, expression of His-tagged enzyme in Escherichia coli strain Rosetta (DE3) pLys
His-tagged version expressed in Escherichia coli BL21
-
His-tagged version expressed in Escherichia coli BL21(DE3)pLys
-
into the vector pET22b+ for expression in Eschericha coli cells
overexpression in Escherichia coli
-
recombinant His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
K0Y2E6
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
gene npl is downregulated by 17beta-estradiol
gene npl is significantly upregulated by progesterone. Progesterone receptor antagonist RU486 blocks the upregulation of Npl in both preimplantation uterus and progesterone-treated ovariectomized uterus
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D187E
-
increased Km
D187N
-
decreased Vmax, increased Km
E188D
-
cleaves 5-N-acetyl-9-O-acetylneuraminic acid as good as N-acetylneuraminic acid
E188Q
-
cleaves 5-N-acetyl-9-O-acetylneuraminic acid as good as N-acetylneuraminic acid
K161R
-
decreased Vmax
Y133C
-
significant decrease in enzyme activity
Y133F
-
significant decrease in enzyme activity
Y133H
-
significant decrease in enzyme activity
Y133W
-
significant decrease in enzyme activity
D191P
-
ratio of turnover-number to KM-value for (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid is 1.4fold higher than the wild-type ratio
E192A
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192C
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192D
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192F
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192G
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192H
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192I
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192K
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192L
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192M
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192P
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192Q
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192R
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192T
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192W
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192Y
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
-
KM-value for N-acetyl-D-neuraminic acid is 5.9fold higher than the wild-type value, turnover-number for N-acetyl-D-neuraminic acid is 14% of the wild-type value, KM-value for N-acetyl-L-neuraminic acid is 7.6fold higher than the wild-type value, turnover-number for N-acetyl-L-neuraminic acid is 100fold higher than the wild-type value, KM-value for D-3-deoxy-manno-2-octulosonic acid is 83% of the wild-type value, turnover-number for D-3-deoxy-manno-2-octulosonic acid is 7.4fold higher than wild-type value, KM-value for L-3-deoxy-manno-2-octulosonic acid is 37% of the wild-type value, turnover-number for L-3-deoxy-manno-2-octulosonic acid is 9.5fold higher than wild-type value. kcat/KM for N-acetyl-D-neuraminic acid is 40.4fold lower than the wild-type value, kcat/KM for N-acetyl-D-neuraminic acid is 20fold higher than the wild-type value, kcat/KM for L-3-deoxy-manno-2-octulosonic acid is 24.7fold higher than the wild-type value
I229D
the mutant shows decreased kcat compared to the wild type enzyme
I229N
the mutant shows increased kcat compared to the wild type enzyme
I229R
the mutant shows increased kcat compared to the wild type enzyme
L142R
-
increased activity towards L-aspartate-beta-semialdehyde
L142R/Y190D/E192A
-
less efficient than wild-type enzyme
L171D
the mutant is insoluble
L171D/L199D/I229D
the mutant is insoluble
L171D/L199N/I229D
the mutant is insoluble
L171N
the mutant is insoluble
L171N/I229N
the mutant shows decreased kcat compared to the wild type enzyme
L171R
the mutant shows decreased kcat compared to the wild type enzyme
L171R/L199N/I229R
the mutant is insoluble
L171R/L199R/I229R
the mutant is insoluble
L199D
the mutant is insoluble
L199N
the mutant shows increased kcat compared to the wild type enzyme
L199N/I229D
the mutant shows decreased kcat compared to the wild type enzyme
L199N/I229N
the mutant shows decreased kcat compared to the wild type enzyme
L199N/I229R
the mutant shows increased kcat compared to the wild type enzyme
L199R
the mutant is insoluble
L199R/I229N
the mutant shows increased kcat compared to the wild type enzyme
S208G
-
ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 2.5fold lower than the wild-type ratio, ratio of turnover-number to KM-value for (2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid is 1.5fold higher than the wild-type ratio
Y98H/F115L
-
KM-value for N-acetyl-D-neuraminic acid is 81% of the wild-type value, turnover-number for N-acetyl-D-neuraminic acid isnearly identical to wild-type value, KM-value for D-3-deoxy-manno-2-octulosonic acid is 89% of the wild-type value, turnover-number for D-3-deoxy-manno-2-octulosonic acid is 1.25fold higher than wild-type value, KM-value for L-3-deoxy-manno-2-octulosonic acid is 1.9fold higher than the wild-type value, turnover-number for L-3-deoxy-manno-2-octulosonic acid is identical to wild-type value. kcat/KM for N-acetyl-D-neuraminic acid is 1.1fold higher than the wild-type ratio, kcat/Km for D-3-deoxy-manno-2-octulosonic acid is 1.4fold higher than the wild-type ratio, kcat/KM for L-3-deoxy-manno-2-octulosonic acid is 1.8fold lower than the wild-type ratio
Y98H/F115L/N153Y/V251I/V265I/Y281C
-
KM-value for N-acetyl-D-neuraminic acid is 3.9fold higher than the wild-type value, turnover-number for N-acetyl-D-neuraminic acid is 16% of the wild-type value, KM-value for N-acetyl-L-neuraminic acid is 2.6fold higher than the wild-type value, turnover-number for N-acetyl-L-neuraminic acid is 100fold higher than the wild-type value, KM-value for D-3-deoxy-manno-2-octulosonic acid is 46% of the wild-type value, turnover-number for D-3-deoxy-manno-2-octulosonic acid is 4.36fold higher than wild-type value, KM-value for L-3-deoxy-manno-2-octulosonic acid is 32% of the wild-type value, turnover-number for L-3-deoxy-manno-2-octulosonic acid is 3.8fold higher than wild-type value. kcat/Km for N-acetyl-D-neuraminic acid is 23.8fold lower than the wild-type value, kcat/Km for N-acetyl-D-neuraminic acid is 20fold higher than the wild-type value, kcat/Km for D-3-deoxy-manno-2-octulosonic acid is 9.3fold higher than the wild-type value
Y98H/F115L/V251I
-
KM-value for N-acetyl-D-neuraminic acid is 1.5fold higher than the wild-type value, turnover-number for N-acetyl-D-neuraminic acid is 92% of the wild-type value, KM-value for N-acetyl-L-neuraminic acid is 84fold higher than the wild-type value, turnover-number for N-acetyl-L-neuraminic acid is 22.5fold higher than the wild-type value, KM-value for D-3-deoxy-manno-2-octulosonic acid is nearly identical to the wild-type value, turnover-number for D-3-deoxy-manno-2-octulosonic acid is 2.3fold higher than wild-type value, KM-value for L-3-deoxy-manno-2-octulosonic acid is 1.2fold higher than the wild-type value, turnover-number for L-3-deoxy-manno-2-octulosonic acid is 1.7fold higher than wild-type value. kcat/KM for N-acetyl-D-neuraminic acid is 1.6fold lower than the wild-type value, kcat/KM for N-acetyl-D-neuraminic acid is 5fold lower than the wild-type value, kcat/KM for D-3-deoxy-manno-2-octulosonic acid is 2.2fold higher than the wild-type value, kcat/KM for L-3-deoxy-manno-2-octulosonic acid is 1.4fold higher than the wild-type value
Y98H/F115L/V251I/V265I
-
KM-value for N-acetyl-D-neuraminic acid is 4.5fold higher than the wild-type value, turnover-number for N-acetyl-D-neuraminic acid is 45% of the wild-type value, KM-value for N-acetyl-L-neuraminic acid is 28.9fold higher than the wild-type value, turnover-number for N-acetyl-L-neuraminic acid is 34fold higher than the wild-type value, KM-value for D-3-deoxy-manno-2-octulosonic acid is 25% of the wild-type value, turnover-number for D-3-deoxy-manno-2-octulosonic acid is 2.6fold higher than wild-type value, KM-value for L-3-deoxy-manno-2-octulosonic acid is 36% of the wild-type value, turnover-number for L-3-deoxy-manno-2-octulosonic acid is 2.3fold higher than wild-type value. kcat/Km for N-acetyl-D-neuraminic acid is 22.4fold lower than the wild-type value, kcat/Km for N-acetyl-D-neuraminic acid is 2fold higher than the wild-type value
L171N/I229N
-
the mutant shows decreased kcat compared to the wild type enzyme
-
L199N/I229D
-
the mutant shows decreased kcat compared to the wild type enzyme
-
L199N/I229N
-
the mutant shows decreased kcat compared to the wild type enzyme
-
L199N/I229R
-
the mutant shows increased kcat compared to the wild type enzyme
-
L199R/I229N
-
the mutant shows increased kcat compared to the wild type enzyme
-
E192N
-
the turnover-number for sialic acid is 65% of the wild-type value, the Km-value for sialic acid is 8.6fold higher than the wild-type value, the turnover-number for (2R,3R,4R)-6-[(aminooxy)carbonyl]-3,4,6-trihydroxy-N-propyltetrahydro-2H-pyran-2-carboxamide is 57% of the wild-type value, the KM-value for (2R,3R,4R)-6-[(aminooxy)carbonyl]-3,4,6-trihydroxy-N-propyltetrahydro-2H-pyran-2-carboxamide is 28.2fold lower than the wild-type value. The mutant enzyme is most efficient in catalysis of the synthesis of the tertiary amides (4S,5R,6R)-6-[(diethylamino)carbonyl]-2,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid, (4R,5R,6R)-6-[(diethylamino)carbonyl]-2,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid, (4S,5R,6R)-2,4,5-trihydroxy-6-{[methyl(propyl)amino]carbonyl}tetrahydro-2H-pyran-2-carboxylic acid, (4R,5R,6R)-2,4,5-trihydroxy-6-{[methyl(propyl)amino]carbonyl}tetrahydro-2H-pyran-2-carboxylic acid, (4S,5R,6R)-6-[(dipropylamino)carbonyl]-2,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid, (4R,5R,6R)-6-[(dipropylamino)carbonyl]-2,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid, (4S,5R,6R)-6-[(dibutylamino)carbonyl]-2,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid, (4R,5R,6R)-6-[(dibutylamino)carbonyl]-2,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid, (4S,5R,6R)-2,4,5-trihydroxy-6-(pyrrolidin-1-ylcarbonyl)tetrahydro-2H-pyran-2-carboxylic acid, (4R,5R,6R)-2,4,5-trihydroxy-6-(pyrrolidin-1-ylcarbonyl)tetrahydro-2H-pyran-2-carboxylic acid, (4S,5R,6R)-2,4,5-trihydroxy-6-(piperidin-1-ylcarbonyl)tetrahydro-2H-pyran-2-carboxylic acid, (4R,5R,6R)-2,4,5-trihydroxy-6-(piperidin-1-ylcarbonyl)tetrahydro-2H-pyran-2-carboxylic acid, (4S,5R,6R)-2,4,5-trihydroxy-6-(morpholin-4-ylcarbonyl)tetrahydro-2H-pyran-2-carboxylic acid, (4R,5R,6R)-2,4,5-trihydroxy-6-(morpholin-4-ylcarbonyl)tetrahydro-2H-pyran-2-carboxylic acid
F252A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F252Y
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S47A
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
S47C
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
S47T
-
site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme
T167A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T167S
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
T48A
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
T48S
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
Y110A
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Y110F
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y137A
-
site-directed mutagenesis, inactive mutant
Y137F
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
K164A
K0Y2E6
site-directed mutagenesis, determiniation of three structures of the K164A variant: one in the absence of substrates and two binary complexes with N-acetylneuraminic acid and N-glycolylneuraminic acid. Both sialic acids bind to the active site in the open-chain ketone form of the monosaccharide, every hydroxyl group of the linear sugars makes hydrogen bond interactions with the enzyme
C118A
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
C118S
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
C238A
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
C238S
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
C270A
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
C270S
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
C82A
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
C82S
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
K165C
site-directed mutagenesis to introduce a cysteine in place of Lys165 in the enzyme active site and complete conversion of the cysteine into gamma-thialysine through dehydroalanine as by chemical mutagenesis, ESI-mass spectrometry and kinetic characterisation, the K165C variant is severely impaired in catalysis, kcat/Km is reduced 720fold compared with wild-type
K165Dha
site-directed mutagenesis to introduce a cysteine in place of Lys165 in the enzyme active site and complete conversion of the cysteine into gamma-thialysine, Dha, through dehydroalanine as by chemical mutagenesis, ESI-mass spectrometry and kinetic characterisation, the enzyme containing gamma-thialysine regains 17-30% of the wild-type activity
C238S
-
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
-
C270S
-
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
-
C82A
-
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
-
C82S
-
site-directed mutagenesis, the mutant shows kinetic properties identical to the wild-type enzyme
-
K165C
-
site-directed mutagenesis to introduce a cysteine in place of Lys165 in the enzyme active site and complete conversion of the cysteine into gamma-thialysine through dehydroalanine as by chemical mutagenesis, ESI-mass spectrometry and kinetic characterisation, the K165C variant is severely impaired in catalysis, kcat/Km is reduced 720fold compared with wild-type
-
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type enzyme with various concentrations of urea in Tris·HCl buffer, 50 mM, pH 7.4, the refolded enzyme shows higher activity than the native wild-type enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
synthesis
Show AA Sequence (1508 entries)
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