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Information on EC 4.1.3.3 - N-acetylneuraminate lyase and Organism(s) Escherichia coli and UniProt Accession P0A6L4
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4.1.3.3 N-acetylneuraminate lyaseIUBMB Comments
This enzyme is involved in the degradation of N-acetylneuraminate. It is specific for the open form of the sugar. It also acts on N-glycoloylneuraminate and on O-acetylated sialic acids, other than 4-O-acetylated derivatives.
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Word Map
- 4.1.3.3
- mannac
-
2-epimerase
- n-acetylmannosamine
- aldolases
-
n-glycolylneuraminic
-
cmp-sialic
- dihydrodipicolinate
-
3.2.1.18
- biotechnology
- neu5gc
- dhdps
- synthesis
- n-acylneuraminate
- analysis
- industry
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
n-acetylneuraminate lyase, sialic acid aldolase, n-acetylneuraminic acid aldolase, n-acetylneuraminic acid lyase, neu5ac aldolase, n-acetyl-d-neuraminic acid aldolase, neuac lyase, n-acetylneuraminate pyruvate lyase, n-acetylneuraminate pyruvate-lyase, neuraminic acid aldolase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Acetylneuraminate pyruvate-lyase
-
-
-
-
Lyase, acetylneuraminate
-
-
-
-
N-acetyl-D-neuraminic acid lyase
N-Acetylneuraminate aldolase
-
-
-
-
N-Acetylneuraminate lyase
-
-
-
-
N-acetylneuraminate pyruvate lyase
-
-
-
-
N-Acetylneuraminate pyruvate-lyase
-
-
-
-
N-Acetylneuraminic acid aldolase
N-Acetylneuraminic acid lyase
N-Acetylneuraminic aldolase
-
-
-
-
N-Acetylneuraminic lyase
-
-
-
-
NAL
NALase
-
-
-
-
NANA lyase
-
-
-
-
NeuAc aldolase
-
-
-
-
Neuraminate aldolase
-
-
-
-
Neuraminic acid aldolase
-
-
-
-
Neuraminic aldolase
-
-
-
-
NPL
-
-
-
-
Sialate lyase
-
-
-
-
Sialic acid aldolase
Sialic aldolase
-
-
-
-
N-Acetylneuraminic acid aldolase
-
-
-
-
N-Acetylneuraminic acid lyase
-
-
-
-
Sialic acid aldolase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
condensation
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
aceneuramate pyruvate-lyase (N-acetyl-D-mannosamine-forming)
This enzyme is involved in the degradation of N-acetylneuraminate. It is specific for the open form of the sugar. It also acts on N-glycoloylneuraminate and on O-acetylated sialic acids, other than 4-O-acetylated derivatives.
CAS REGISTRY NUMBER
COMMENTARY
9027-60-5
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
?
-
-
-
-
?
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
pyruvate + (2R,3S)-2,3-dihydroxy-4-oxo-N,N-dipropylbutanamide
-
-
-
-
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
-
-
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
-
-
34% yield
-
?
3-bromo-3-deoxy-D-mannose + pyruvate
?
-
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
-
-
?
4-O-alpha-D-galactopyranosyl-D-mannopyranose + pyruvate
3-deoxy-7-O-alpha-D-galactopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
-
-
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
-
-
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
-
-
35% yield
-
?
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
-
-
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
-
-
85% yield
-
?
6-O-alpha-D-galactopyranosyl-D-mannopyranose + pyruvate
3-deoxy-9-O-alpha-D-galactopyranosyl-D-glycero-beta-D-galacto-non-2-ulopyranosonic acid
-
-
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
-
-
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
-
-
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
-
-
83% yield
-
?
D-arabinose + pyruvate
D-3-deoxy-manno-2-octulosonic acid
-
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
?
-
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
?
-
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
?
-
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
?
-
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
-
-
-
-
?
D-talose + pyruvate
?
-
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
?
-
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-arabinose + pyruvate
L-3-deoxy-manno-2-octulosonic acid
-
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
?
-
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
?
-
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
-
-
?
N-acetyl-D-mannosamine + pyruvate
N-acetyl-D-neuraminic acid
-
-
-
-
r
N-acetyl-L-mannosamine + pyruvate
N-acetyl-L-neuraminic acid
-
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-neuraminic acid
N-acetyl-D-mannosamine + pyruvate
-
-
-
-
r
N-Acetylneuraminic acid
?
-
N-acetylneuraminate lyase plays important role in regulation of N-acetylneuraminic acid metabolism
-
-
?
pyruvate + alpha-D-Galp-(1,6)-D-Manp
alpha-D-Galp-(1,9)-D-deoxyneuraminic acid
-
-
-
-
?
N-acetyl-D-neuraminic acid
N-acetyl-D-mannosamine + pyruvate
-
-
-
-
r
N-Acetylmannosamine + pyruvate
?
-
N-acetylneuraminic acid is intermediate for colomic acid biosynthesis
-
-
?
N-Acetylneuraminic acid
N-Acetyl-D-mannosamine + pyruvate
-
-
?
N-Acetylneuraminic acid
N-Acetyl-D-mannosamine + pyruvate
-
-
r
N-Acetylneuraminic acid
N-Acetyl-D-mannosamine + pyruvate
-
syn: Neu5Ac
-
r
additional information
?
-
-
condensation: N-acetylglucosamine, N-acetylgalactosamine, glucosamine and galactosamine cannot replace N-acetylmannosamine. Phosphoenolpyruvate, lactate, beta-hydroxypyruvate and other pyruvate derivatives cannot replace pyruvate
-
-
?
additional information
?
-
-
no substrates are 3-azido-3-deoxy, 3-amino-3-deoxy and 3-N-tert-butylcarbonyl-3-deoxy derivatives of 2-acetamido-2-deoxy-alpha,beta-D-mannose
-
?
additional information
?
-
One point mutation (L142R) is sufficient to create an enzyme that can complement a bacterial auxotroph lacking the gene for dihydrodipicolinate synthase, EC 4.2.1.52. The mutant enzyme catalyzes the reaction: L-aspartate 4-semialdehyde + pyruvate = dihydrodipicolinate + 2 H2O
-
?
additional information
?
-
-
One point mutation (L142R) is sufficient to create an enzyme that can complement a bacterial auxotroph lacking the gene for dihydrodipicolinate synthase, EC 4.2.1.52. The mutant enzyme catalyzes the reaction: L-aspartate 4-semialdehyde + pyruvate = dihydrodipicolinate + 2 H2O
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
N-Acetylneuraminic acid
?
-
N-acetylneuraminate lyase plays important role in regulation of N-acetylneuraminic acid metabolism
-
-
?
N-Acetylmannosamine + pyruvate
?
-
N-acetylneuraminic acid is intermediate for colomic acid biosynthesis
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
hydrogen peroxide
-
0% residual activity at 10 mM, 16% residual activity at 1 mM
L-lysine
50% inhibition at 0.2 mM, inhibits wild-type enzyme, but not mutant enzymes
N-bromosuccinimide
-
0% residual activity at 0.1 mM, 36% residual activity at 0.01 mM
N-bromosuccinimide
-
0% residual activity at 0.01 mM, 36% residual activity at 0.001 mM
p-chloromercuribenzoate
-
0% residual activity at 0.01 mM, 44% residual activity at 0.001 mM
p-chloromercuribenzoate
-
0% residual activity at 0.1 mM, 9% residual activity at 0.01 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.35 - 10.6
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
0.35
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme E192V
0.39
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme E192N
0.5
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme E192S
2.54
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme S208G
5.7
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme D191P
10.6
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, wild-type enzyme
0.2
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192M mutant protein, pH 7.5, 25°C
0.2
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192P mutant protein, pH 7.5, 25°C
0.3
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192F mutant protein, pH 7.5, 25°C
0.4
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192N mutant protein, pH 7.7.5, 25°C
0.4
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192V mutant protein, pH 7.5, 25°C
0.5
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192H mutant protein, pH 7.5, 25°C
0.6
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192S mutant protein, pH 7.5, 25°C
0.7
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192Q mutant protein, pH 7.5, 25°C
1.1
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192D mutant protein, pH 7.5, 25°C
11
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
wild type (E192), pH 7.5, 25°C
3.5
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
6.3
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
11.4
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
12.3
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L
14.5
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
2.4
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
2.7
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
2.8
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
9.2
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
14.1
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L
3.9
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
10.1
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
12.1
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
15.3
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
3 - 3.5
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
36.2
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
127
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
369.4
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
2.51
N-acetylneuraminate
mutant enzyme L171N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
3.2
N-acetylneuraminate
mutant enzyme L199N/I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
3.4
N-acetylneuraminate
mutant enzyme I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
3.42
N-acetylneuraminate
mutant enzyme L171R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
3.93
N-acetylneuraminate
mutant enzyme I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
4
N-acetylneuraminate
mutant enzyme L199N/I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
4.1
N-acetylneuraminate
wild type enzyme, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
4.3
N-acetylneuraminate
mutant enzyme L199N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
4.7
N-acetylneuraminate
mutant enzyme L199N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
5
N-acetylneuraminate
mutant enzyme I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
6.1
N-acetylneuraminate
mutant enzyme L199R/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.44 - 2.18
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
0.44
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme S208G
0.91
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme D191P
1.22
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, wild-type enzyme
1.58
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme E192V
1.85
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme E192S
2.18
(2S,4S,5R,6R)-5-(acetylamino)-6-[(dipropylamino)carbonyl]-2,4-dihydroxytetrahydro-pyran-2-carboxylic acid
-
pH 7.5, 25°C, mutant enzyme E192N
0.1
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192D mutant protein, pH 7.5, 25°C
0.43
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192S mutant protein, pH 7.5, 25°C
0.63
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192H mutant protein, pH 7.5, 25°C
0.82
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192P mutant protein, pH 7.5, 25°C
1.17
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192M mutant protein, pH 7.5, 25°C
1.22
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
wild type (E192), pH 7.5, 25°C
1.47
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192F mutant protein, pH 7.5, 25°C
1.58
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192V mutant protein, pH 7.5, 25°C
2.17
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192N mutant protein, pH 7.7.5, 25°C
2.83
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192Q mutant protein, pH 7.5, 25°C
1.5
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L
2.8
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
3.2
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
5.1
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
8.9
D-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
1.1
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L
1.9
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
2.5
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
4.2
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
10.4
L-3-deoxy-manno-2-octulosonic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
1.5
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
1.7
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
2.2
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
9.7
N-acetyl-D-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
0.0045
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I
0.0068
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/V251I/V265I
0.02
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme E60A/Y98H/F115L/N153Y/D150G/V251I/V265I/Y281C/N153Y
0.02
N-acetyl-L-neuraminic acid
-
pH 7.5, 25°C, mutant enzyme Y98H/F115L/N153Y/V251I/V265I/Y281C
0.29
N-acetylneuraminate
mutant enzyme L199N/I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
0.664
N-acetylneuraminate
mutant enzyme L199N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
3.15
N-acetylneuraminate
mutant enzyme L171N/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
4.69
N-acetylneuraminate
mutant enzyme I229D, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
5.49
N-acetylneuraminate
mutant enzyme L171R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
6.75
N-acetylneuraminate
wild type enzyme, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
7.4
N-acetylneuraminate
mutant enzyme I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
7.72
N-acetylneuraminate
mutant enzyme I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
7.94
N-acetylneuraminate
mutant enzyme L199N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
8.9
N-acetylneuraminate
mutant enzyme L199N/I229R, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
11.87
N-acetylneuraminate
mutant enzyme L199R/I229N, in sodium phosphate buffer (100 mM, pH 7.6), at 30°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.08
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192D mutant protein, pH 7.5, 25°C
0.12
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
wild type (E192), pH 7.5, 25°C
0.7
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192S mutant protein, pH 7.5, 25°C
1.33
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192H mutant protein, pH 7.5, 25°C
4.17
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192Q mutant protein, pH 7.5, 25°C
4.33
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192P mutant protein, pH 7.5, 25°C
4.5
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192V mutant protein, pH 7.5, 25°C
5.17
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192F mutant protein, pH 7.5, 25°C
5.33
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192M mutant protein, pH 7.5, 25°C
5.67
(5R,6R)-7-(dipropylamino)-4,5,6-trihydroxy-2,7-dioxoheptanoic acid
-
E192N mutant protein, pH 7.7.5, 25°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.3
(2R,3R)-2,3,4-trihydroxy-N,N-dipropylbutanamide
-
E192N mutant protein , pH 7.5, 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
36.26
-
after 14.8fold purification, in 50 mM potassium phosphate buffer, pH 7.5, at 37°C
additional information
additional information
-
after a 36 h incubation, the concentrations of N-acetyl-D-neuraminic acid from the reactions utilizing 0.4, 0.8, 1.2, 1.6, 2.0, and 2.2 M pyruvate as the carbon source are 7.5 (3.75%), 26.9 (3.45%), 43.9 (21.95%), 56.0 (28%), and 61.9 mM (30.95%), respectively. When using 2.2 M pyruvate, in the first 6 h, the concentration of N-acetyl-D-neuraminic acid reaches 29.5 mM (which is about half of the maximal concentration), and the productivity of N-acetyl-D-neuraminic acid is 1.6 g/l/h, the concentration of the intermediate N-acetyl-D-mannosamine also increases gradually from 0 to 64 mM during the process
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
the quaternary structure of Escherichia coli N-acetylneuraminate lyase is essential for functional expression
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
33000
35000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
tetramer
trimer
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
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
E192N
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
E192S
E192T
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192V
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
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
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/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
additional information
E192N
-
ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 13.4fold 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 48.7fold higher than the wild-type ratio
E192N
the mutant has a 6fold higher specificity (kcat/Km) for dipropylaminocarbonyl-substituted derivatives than wild type enzyme
E192N
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192S
-
ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 3.9fold 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 32.2fold higher than the wild-type ratio
E192S
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
E192V
-
ratio of turnover-number to KM-value for N-acetyl-D-neuraminic acid is 17.9fold 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 39.3fold higher than the wild-type ratio
E192V
-
strong interaction of Glu192 (wild type) with hydroxyl groups at C8 and C9 of N-acetylneuraminic acid
additional information
-
directed evolution of D-sialic acid aldolase to L-3-deoxy-manno-2-octulosonic acid (L-KDO) aldolase
additional information
-
synthesis of N-acetylneuraminic acid from N-acetylglucosamine and pyruvate by constructing and co-expressing a polycistronic plasmid encoding an N-acetylglucosamine 2-epimerase gene from Synechocystis sp. PCC 6803 (snAGE) or Anabaena sp. CH1 (anAGE) and the Escherichia coli N-acetylneuraminic acid lyase gene. Construction of four polycistronic plasmids in which the positions of AGE gene differ with respect to Nal gene, plasmid pET-28a-Nal-anAGE with anAGE gene located next to Nal gene produces the highest amount of N-acetylneuraminic acid, overview. anAGE from Anabaena sp. CH1 shows much higher activity and productivity than snAGE from Synechocystis sp. PCC 6803
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
LITERATURE
37
80
additional information
37
-
in 50 mM Tris/HCl, pH 8, 25 mM MgCl2, 1 mM PMSF stable for at least 24 h
80
-
77% loss of activity after 10 min, in presence of 5 mM pyruvate 8% loss of activity after 10 min
additional information
-
thermostability of enzyme produced by recombinant Serratia liquefaciens is higher by about 10°C than thermostability of enzyme produced by recombinant E. coli
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
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
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
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
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
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a Bacillus subtilis outer spore coat protein fusion expressed in Bacillus subtilis WB600 results in a spore-displayed N-acetyl-D-neuraminic acid aldolase
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expressed in Bright Yellow 2 (BY2) tobacco cells and Medicago sativa cytosol
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expressed in Escherichia coli BL21(DE3) cells
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)
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
synthesis
biotechnology
-
efficient method for N-acetyl-D-neuraminic acid production using coupled bacterial cells with a safe temperature-induced system, a precursor for producing many pharmaceutical drugs such as zanamivir which have been used in clinical trials to treat and prevent the infection with influenza virus, such as the avian influenza virus H5N1 and the current 2009 H1N1
biotechnology
-
chemoenzymatic synthesis of N-acetyl-D-neuraminic acid from N-acetyl-D-glucosamine and pyruvate
synthesis
-
enzymatic synthesis of N-acetylneuraminic acid with immobilized enzyme
synthesis
-
enzymatic synthesis of N-acetylneuraminic acid, large scale production using N-acetylneuraminate lyase and N-acetyl-D-glucosamine-2-epimerase
synthesis
-
production of N-acetyl-D-neuraminic acid, which is the major representative of amino sugars. The synthesis of N-acetyl-D-neuraminic acid is of interest in studies towards inhibitors of neuraminidase, hemagglutinin and selectin-mediated leucocyte adhesion
synthesis
-
method to produce N-acetylneuraminic acid efficiently. Using a recombinant human renin binding protein (rhRnBp) showing GlcNAc-2-epimerase activity and Escherichia coli sialic acid aldolase, about 80% conversion yield of Neu5Ac is obtained in the coupling reaction under 10fold excess of pyruvate to GlcNAc based on the initial concentration of GlcNAc. The equilibrium of GlcNAc-2-epimerase reaction is not affected by temperature, whereas that of sialic acid aldolase reaction is shifted toward Neu5Ac by lowering the reaction temperature. Low temperatures improve the conversion yield of Neu5Ac, but decrease the reaction rate in the coupling reaction. A high reaction rate as well as a high conversion yield can be achieved by shifting the temperature of the coupling reaction during the reaction
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Uchida, Y.; Tsukada, Y.; Sugimori, T.
Purification and properties of N-acetylneuraminate lyase from Escherichia coli
J. Biochem.
96
507-522
1984
Escherichia coli
Gross, H.J.; Brossmer, R.
Inhibition of N-acetylneuraminate lyase by N-acetyl-4-oxo-D-neuraminic acid
FEBS Lett.
232
145-147
1988
Clostridium perfringens, Escherichia coli
Aisaka, K.; Tamura, S.; Arai, Y.; Uwajima, T.
Hyperproduction of N-acetylneuraminate lyase by the gene-cloned strain of Escherichia coli
Biotechnol. Lett.
9
633-637
1987
Escherichia coli
-
Ohta, Y.; Shimosaka, M.; Murata, K.; Tsukada, Y.; Kimura, A.
Molecular cloning of the N-acetylneuraminate lyase gene in Escherichia coli K-12
Appl. Microbiol. Biotechnol.
24
386-391
1986
Escherichia coli
-
Comb, D.G.; Roseman, S.
The sialic acids. I. Structure and enzymatic synthesis of N-acetylneuraminic acid
J. Biol. Chem.
235
2529-2537
1960
Escherichia coli, Rattus norvegicus
Yamamoto, K.; Kawakami, B.; Kawamura, Y.; Kawai, K.
Serratia liquefaciens as a new host superior for overproduction and purification using the N-acetylneuraminate lyase gene of Escherichia coli
Anal. Biochem.
246
171-175
1997
Escherichia coli
Aisaka, K.; Igarashi, A.; Yamaguchi, K.; Uwajima, T.
Purification, crystallization and characterization of N-acetylneuraminate lyase from Escherichia coli
Biochem. J.
276
541-546
1991
Escherichia coli
Izard, T.; Lawrence, M.C.; Malby, R.L.; Lilley, G.G.; Colman, P.M.
The three-dimensional structure of N-acetylneuraminate lyase from Escherichia coli
Structure
2
361-369
1994
Escherichia coli
Maru, I.; Ohnishi, J.; Ohta, Y.; Tsukada, Y.
Simple and large-scale production of N-acetylneuraminic acid from N-acetyl-D-glucosamine and pyruvate using N-acyl-D-glucosamine 2-epimerase and N-acetylneuraminate lyase
Carbohydr. Res.
306
575-578
1998
Escherichia coli
Mahmoudian, M.; Noble, D.; Drake, C.S.; Middleton, R.F.; Montgomery, D.S.; Piercey, J.E.; Ralakhan, D.; Todd, M.; Dawson, M.J.
An efficient process for production of N-acetylneuraminic acid using N-acetylneuraminic acid aldolase
Enzyme Microb. Technol.
20
393-400
1997
Escherichia coli
Lilley, G.G.; Von Itzstein, M.; Ivancic, N.
High-level production and purification of Escherichia coli N-acetylneuraminic acid aldolase (EC 4.1.3.3)
Protein Expr. Purif.
3
434-440
1992
Escherichia coli
Ferrero, M.A.; Reglero, A.; Fernandez-Lopez, M.; Ordas,.; Rodriguez-Aparicio, L.B.
N-acetyl-D-neuraminic acid lyase generates the sialic acid for colominic acid biosynthesis in Escherichia coli K1
Biochem. J.
317
157-165
1996
Escherichia coli
Wada, M.; Hsu, C.C.; Franke, D.; Mitchell, M.; Heine, A.; Wilson, I.; Wong, C.H.
Directed evolution of N-acetylneuraminic acid aldolase to catalyze enantiomeric aldol reactions
Bioorg. Med. Chem.
11
2091-2098
2003
Escherichia coli (P0A6L4), Escherichia coli
Kok, G.B.; Campbell, M.; Mackey, B.L.; von Itzstein, M.
Synthesis of C-3 nitrogen-containing derivatives of N-acetyl-alpha,beta-D-mannosamine as substrates for N-acetylneuraminic acid aldolase
Carbohydr. Res.
332
133-139
2001
Escherichia coli
Joerger, A.C.; Mayer, S.; Fersht, A.R.
Mimicking natural evolution in vitro: an N-acetylneuraminate lyase mutant with an increased dihydrodipicolinate synthase activity
Proc. Natl. Acad. Sci. USA
100
5694-5699
2003
Escherichia coli (P0A6L4), Escherichia coli
Mahmoudian, M.; Noble, D.; Drake, C.S.; Middleton, R.F.; Montgomery, D.S.; Piercey, J.E.; Ramlakhan, D.; Todd, M.; Dawson, M.J.
An efficient process for production of N-acetylneuraminic acid using N-acetylneuramic acid aldolase
Enzyme Microb. Technol.
45
393-400
1997
Escherichia coli
-
Lee, J.O.; Yi, J.K.; Lee, S.G.; Takahashi, S.; Kim, B.G.
Production of N-acetylneuraminic acid from N-acetylglucosamine and pyruvate using recombinant human renin binding protein and sialic acid aldolase in one pot
Enzyme Microb. Technol.
35
121-125
2004
Escherichia coli
-
Hsu, C.C.; Hong, Z.; Wada, M.; Franke, D.; Wong, C.H.
Directed evolution of D-sialic acid aldolase to L-3-deoxy-manno-2-octulosonic acid (L-KDO) aldolase
Proc. Natl. Acad. Sci. USA
102
9122-9126
2005
Escherichia coli
Williams, G.J.; Woodhall, T.; Nelson, A.; Berry, A.
Structure-guided saturation mutagenesis of N-acetylneuraminic acid lyase for the synthesis of sialic acid mimetics
Protein Eng. Des. Sel.
18
239-246
2005
Escherichia coli
Lee, Y.C.; Chien, H.C.; Hsu, W.H.
Production of N-acetyl-D-neuraminic acid by recombinant whole cells expressing Anabaena sp. CH1 N-acetyl-D-glucosamine 2-epimerase and Escherichia coli N-acetyl-D-neuraminic acid lyase
J. Biotechnol.
129
453-460
2007
Escherichia coli
Huang, S.; Yu, H.; Chen, X.
Disaccharides as sialic acid aldolase substrates: Synthesis of disaccharides containing a sialic acid at the reducing end
Angew. Chem. Int. Ed. Engl.
46
2249-2253
2007
Escherichia coli
Wang, T.; Lee, W.
Production of 2-keto-3-deoxy-D-glycero-D-galacto-nonopyranulosonic acid (KDN) using fusion protein of N-acetyl-D-neuraminic acid aldolase
Biochem. Eng. J.
29
75-80
2006
Escherichia coli
-
Yu, H.; Chen, X.
Aldolase-catalyzed synthesis of beta-D-Galp-(1-->9)-D-KDN: a novel acceptor for sialyltransferases
Org. Lett.
8
2393-2396
2006
Escherichia coli
Paccalet, T.; Bardor, M.; Rihouey, C.; Delmas, F.; Chevalier, C.; DAoust, M.A.; Faye, L.; Vezina, L.; Gomord, V.; Lerouge, P.
Engineering of a sialic acid synthesis pathway in transgenic plants by expression of bacterial Neu5Ac-synthesizing enzymes
Plant Biotechnol. J.
5
16-25
2007
Escherichia coli, Escherichia coli K1
Campeotto, I.; Carr, S.B.; Trinh, C.H.; Nelson, A.S.; Berry, A.; Phillips, S.E.; Pearson, A.R.
Structure of an Escherichia coli N-acetyl-D-neuraminic acid lyase mutant, E192N, in complex with pyruvate at 1.45 A resolution
Acta Crystallogr. Sect. F
65
1088-1090
2009
Escherichia coli (P0A6L4), Escherichia coli
Hu, S.; Chen, J.; Yang, Z.; Shao, L.; Bai, H.; Luo, J.; Jiang, W.; Yang, Y.
Coupled bioconversion for preparation of N-acetyl-D-neuraminic acid using immobilized N-acetyl-D-glucosamine-2-epimerase and N-acetyl-D-neuraminic acid lyase
Appl. Microbiol. Biotechnol.
85
1383-1391
2010
Escherichia coli, Sus scrofa (Q9BEC7), Escherichia coli TG1
Zhang, Y.; Tao, F.; Du, M.; Ma, C.; Qiu, J.; Gu, L.; He, X.; Xu, P.
An efficient method for N-acetyl-D-neuraminic acid production using coupled bacterial cells with a safe temperature-induced system
Appl. Microbiol. Biotechnol.
86
481-489
2010
Escherichia coli
Devenish, S.R.; Gerrard, J.A.
The quaternary structure of Escherichia coli N-acetylneuraminate lyase is essential for functional expression
Biochem. Biophys. Res. Commun.
388
107-111
2009
Escherichia coli (P0A6L4), Escherichia coli, Escherichia coli W3110 / ATCC 27325 (P0A6L4)
Gao, C.; Xu, X.; Zhang, X.; Che, B.; Ma, C.; Qiu, J.; Tao, F.; Xu, P.
Chemoenzymatic synthesis of N-acetyl-D-neuraminic acid from N-acetyl-D-glucosamine using the spore surface displayed N-acetyl-D-neuraminic acid aldolase
Appl. Environ. Microbiol.
77
7080-7083
2011
Escherichia coli
Campeotto, I.; Bolt, A.H.; Harman, T.A.; Dennis, C.; Trinh, C.H.; Phillips, S.E.; Nelson, A.; Pearson, A.R.; Berry, A.
Structural insights into substrate specificity in variants of N-acetylneuraminic acid lyase produced by directed evolution
J. Mol. Biol.
404
56-69
2010
Escherichia coli
Sun, W.; Ji, W.; Li, N.; Tong, P.; Cheng, J.; He, Y.; Chen, Y.; Chen, X.; Wu, J.; Ouyang, P.; Xie, J.; Ying, H.
Construction and expression of a polycistronic plasmid encoding N-acetylglucosamine 2-epimerase and N-acetylneuraminic acid lyase simultaneously for production of N-acetylneuraminic acid
Biores. Technol.
130
23-29
2013
Escherichia coli
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