BRENDA - Enzyme Database show
show all sequences of 2.4.99.4

Decreasing the sialidase activity of multifunctional Pasteurella multocida alpha2-3-sialyltransferase 1 (PmST1) by site-directed mutagenesis

Sugiarto, G.; Lau, K.; Li, Y.; Khedri, Z.; Yu, H.; Le, D.T.; Chen, X.; Mol. Biosyst. 7, 3021-3027 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli BL21 (DE3) cells
Pasteurella multocida
Engineering
Amino acid exchange
Commentary
Organism
E271F
the mutation does not affect alpha2-3-sialyltransferase activity
Pasteurella multocida
E271F/R313Y
the mutations do not affect alpha2-3-sialyltransferase activity
Pasteurella multocida
R313Y
the mutation does not affect alpha2-3-sialyltransferase activity
Pasteurella multocida
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.18
-
CMP-N-acetylneuraminate
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.34
-
CMP-N-acetylneuraminate
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.44
-
CMP-N-acetylneuraminate
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.54
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.62
-
CMP-N-acetylneuraminate
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.67
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.71
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
1.4
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pasteurella multocida
-
-
-
Purification (Commentary)
Commentary
Organism
Ni2+-NTA column chromatography
Pasteurella multocida
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
CMP-N-acetylneuraminate + 4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
best substrate, about 37% conversion
723156
Pasteurella multocida
CMP + 4-methylumbelliferyl alpha-N-acetylneuraminyl-(2->3)-beta-D-galactosyl-(1->4)-beta-D-glucoside
-
-
-
-
CMP-N-acetylneuraminate + lactose
-
723156
Pasteurella multocida
CMP + NeuAcalpha(2->3)Galbeta(1->4)Glc
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2
8
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
17
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
19
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
19
-
CMP-N-acetylneuraminate
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
23
-
CMP-N-acetylneuraminate
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
26
-
CMP-N-acetylneuraminate
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
32
-
CMP-N-acetylneuraminate
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
47
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6
10
-
Pasteurella multocida
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21 (DE3) cells
Pasteurella multocida
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
E271F
the mutation does not affect alpha2-3-sialyltransferase activity
Pasteurella multocida
E271F/R313Y
the mutations do not affect alpha2-3-sialyltransferase activity
Pasteurella multocida
R313Y
the mutation does not affect alpha2-3-sialyltransferase activity
Pasteurella multocida
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.18
-
CMP-N-acetylneuraminate
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.34
-
CMP-N-acetylneuraminate
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.44
-
CMP-N-acetylneuraminate
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.54
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.62
-
CMP-N-acetylneuraminate
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.67
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
0.71
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
1.4
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
Purification (Commentary) (protein specific)
Commentary
Organism
Ni2+-NTA column chromatography
Pasteurella multocida
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
CMP-N-acetylneuraminate + 4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
best substrate, about 37% conversion
723156
Pasteurella multocida
CMP + 4-methylumbelliferyl alpha-N-acetylneuraminyl-(2->3)-beta-D-galactosyl-(1->4)-beta-D-glucoside
-
-
-
-
CMP-N-acetylneuraminate + lactose
-
723156
Pasteurella multocida
CMP + NeuAcalpha(2->3)Galbeta(1->4)Glc
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2
8
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
17
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
19
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
19
-
CMP-N-acetylneuraminate
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
23
-
CMP-N-acetylneuraminate
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
26
-
CMP-N-acetylneuraminate
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
32
-
CMP-N-acetylneuraminate
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
47
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6
10
-
Pasteurella multocida
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
28
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
30
-
CMP-N-acetylneuraminate
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
32
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
34
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
39
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
69
-
CMP-N-acetylneuraminate
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
73
-
CMP-N-acetylneuraminate
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
140
-
CMP-N-acetylneuraminate
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
28
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
30
-
CMP-N-acetylneuraminate
mutant enzyme R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
32
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
34
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
39
-
4-methylumbelliferyl 4-O-beta-D-galactosyl-beta-D-glucoside
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
69
-
CMP-N-acetylneuraminate
mutant enzyme E271F/R313Y, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
73
-
CMP-N-acetylneuraminate
wild type enzyme, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
140
-
CMP-N-acetylneuraminate
mutant enzyme E271F, in Tris-HCl buffer (100 mM, pH 8.5), at 37C
Pasteurella multocida
Other publictions for EC 2.4.99.4
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
736507
Lu
Expression of N-acetylglucosam ...
Homo sapiens
J. Biol. Chem.
291
5708-5720
2016
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723156
Sugiarto
Decreasing the sialidase activ ...
Pasteurella multocida
Mol. Biosyst.
7
3021-3027
2011
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1
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3
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8
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1
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1
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2
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8
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1
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1
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3
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8
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1
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2
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8
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1
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8
8
723519
Perez-Garay
alpha2,3-Sialyltransferase ST3 ...
Homo sapiens
PLoS ONE
5
e12524
2010
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2
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1
1
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705677
Huang
Human beta-galactoside alpha-2 ...
Homo sapiens
Mol. Cell. Biochem.
331
81-88
2009
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1
1
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2
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2
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1
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2
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1
1
1
1
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744308
Chandrasekaran
Reversible sialylation synthe ...
Rattus norvegicus
Biochemistry
47
320-330
2008
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1
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1
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2
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746526
Sengupta
-
Chemoenzymatic synthesis of s ...
Rattus norvegicus
Tetrahedron Lett.
44
6037-6042
2003
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1
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744624
Shigeta
alpha-(2->3)- and alpha-(2->6 ...
Mus musculus
Carbohydr. Res.
264
111-121
1994
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745255
Higgins
Aberrant O-linked oligosaccha ...
Homo sapiens
J. Biol. Chem.
266
6280-6290
1991
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745253
Westcott
Reconstitution of a porcine s ...
Sus scrofa
J. Biol. Chem.
260
13116-13121
1985
3
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3
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1
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2
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1
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745248
Sadler
Purification to homogeneity o ...
Sus scrofa
J. Biol. Chem.
254
4434-4443
1979
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3
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1
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1
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2
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1
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2
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1
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745249
Rearick
Enzymatic characterization of ...
Sus scrofa
J. Biol. Chem.
254
4444-4451
1979
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4
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4
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4
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7
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1
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