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Literature summary for 3.2.1.18 extracted from

  • Park, K.H.; Kim, M.G.; Ahn, H.J.; Lee, D.H.; Kim, J.H.; Kim, Y.W.; Woo, E.J.
    Structural and biochemical characterization of the broad substrate specificity of Bacteroides thetaiotaomicron commensal sialidase (2013), Biochim. Biophys. Acta, 1834, 1510-1519.
    View publication on PubMed
Search for more literature for 3.2.1.18

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Bacteroides thetaiotaomicron

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapor diffusion method, using 20% (w/v) polyethylene glycol 4000, 0.2 M ammonium citrate tri-basic (pH 7.0) and 0.1 M Tris pH 8.5 Bacteroides thetaiotaomicron

Inhibitors

Inhibitors Comment Organism Structure
2-deoxy-2,3-dehydro-N-acetyl neuraminic acid competitive inhibition Bacteroides thetaiotaomicron
Cu2+ 2.9% residual activity at 5 mM Bacteroides thetaiotaomicron
Fe2+ 33.7% residual activity at 5 mM Bacteroides thetaiotaomicron
additional information EDTA does not cause a significant decrease in activity Bacteroides thetaiotaomicron
SDS 0.9% residual activity at 5 mM Bacteroides thetaiotaomicron
Zn2+ 31.9% residual activity at 5 mM Bacteroides thetaiotaomicron

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.11
-
 4-nitrophenyl alpha-D-N-acetylneuraminic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
0.16
-
 4-methylumbelliferyl alpha-D-N-acetylneuraminic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
0.29
-
 colominic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
0.49
-
 sialyl-alpha-2,6-lactose pH 7.0 and 37°C Bacteroides thetaiotaomicron
1.47
-
 sialyl-alpha-2,3-lactose pH 7.0 and 37°C Bacteroides thetaiotaomicron

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ the activity is maintained more than 94% in the presence of Ca2+ at concentrations up to 5 mM Bacteroides thetaiotaomicron
Mg2+ the activity is maintained more than 96% in the presence of Mg2+ at concentrations up to 5 mM Bacteroides thetaiotaomicron
Mn2+ the activity is maintained more than 69% in the presence of Mn2+ at concentrations up to 5 mM Bacteroides thetaiotaomicron
Ni2+ the activity is maintained more than 75% in the presence of Ni2+ at concentrations up to 5 mM Bacteroides thetaiotaomicron

Organism

Organism UniProt Comment Textmining
Bacteroides thetaiotaomicron Q8AAK9
-
-
Bacteroides thetaiotaomicron VPI-5482 Q8AAK9
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-affinity column chromatography Bacteroides thetaiotaomicron

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
8.6
-
 with transferrin as substrate, pH 7.0 and 37°C Bacteroides thetaiotaomicron
18.2
-
 with alpha1-acid glycoprotein as substrate, pH 7.0 and 37°C Bacteroides thetaiotaomicron
35
-
 with fetuin as substrate, pH 7.0 and 37°C Bacteroides thetaiotaomicron
64.1
-
 with colominic acid as substrate, pH 7.0 and 37°C Bacteroides thetaiotaomicron
97.7
-
 with sialyl-alpha-2,3-lactose as substrate, pH 7.0 and 37°C Bacteroides thetaiotaomicron
100.9
-
 with sialyl-alpha-2,6-lactose as substrate, pH 7.0 and 37°C Bacteroides thetaiotaomicron

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid + H2O
-
 Bacteroides thetaiotaomicron 4-methylumbelliferone + N-acetylneuraminic acid
-
 ?
4-nitrophenyl alpha-D-N-acetylneuraminic acid + H2O
-
 Bacteroides thetaiotaomicron 4-nitrophenol + N-acetylneuraminic acid
-
 ?
alpha1-acid glycoprotein + H2O
-
 Bacteroides thetaiotaomicron ?
-
 ?
alpha1-acid glycoprotein + H2O
-
 Bacteroides thetaiotaomicron VPI-5482 ?
-
 ?
colominic acid + H2O
-
 Bacteroides thetaiotaomicron ?
-
 ?
colominic acid + H2O
-
 Bacteroides thetaiotaomicron VPI-5482 ?
-
 ?
fetuin + H2O
-
 Bacteroides thetaiotaomicron ?
-
 ?
fetuin + H2O
-
 Bacteroides thetaiotaomicron VPI-5482 ?
-
 ?
sialyl-alpha-2,3-lactose + H2O
-
 Bacteroides thetaiotaomicron ?
-
 ?
sialyl-alpha-2,3-lactose + H2O
-
 Bacteroides thetaiotaomicron VPI-5482 ?
-
 ?
sialyl-alpha-2,6-lactose + H2O
-
 Bacteroides thetaiotaomicron ?
-
 ?
sialyl-alpha-2,6-lactose + H2O
-
 Bacteroides thetaiotaomicron VPI-5482 ?
-
 ?
transferrin + H2O
-
 Bacteroides thetaiotaomicron ?
-
 ?

Synonyms

Synonyms Comment Organism
neuraminidase
-
 Bacteroides thetaiotaomicron
sialidase
-
 Bacteroides thetaiotaomicron

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
 Bacteroides thetaiotaomicron

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 50 more than 60% of activity is maintained from 30 to 50°C Bacteroides thetaiotaomicron

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
82.4
-
 4-nitrophenyl alpha-D-N-acetylneuraminic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
146.7
-
 4-methylumbelliferyl alpha-D-N-acetylneuraminic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
157.6
-
 colominic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
161
-
 sialyl-alpha-2,6-lactose pH 7.0 and 37°C Bacteroides thetaiotaomicron
313.5
-
 sialyl-alpha-2,3-lactose pH 7.0 and 37°C Bacteroides thetaiotaomicron

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 in 50 mM Tris-HCl buffer Bacteroides thetaiotaomicron

pH Range

pH Minimum pH Maximum Comment Organism
6 8 more than 70% of activity is maintained from pH 6.0 to 8.0 Bacteroides thetaiotaomicron

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.035
-
 2-deoxy-2,3-dehydro-N-acetyl neuraminic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
213.3
-
 sialyl-alpha-2,3-lactose pH 7.0 and 37°C Bacteroides thetaiotaomicron
328.6
-
 sialyl-alpha-2,6-lactose pH 7.0 and 37°C Bacteroides thetaiotaomicron
541.2
-
 colominic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
729.2
-
 4-nitrophenyl alpha-D-N-acetylneuraminic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron
916.9
-
 4-methylumbelliferyl alpha-D-N-acetylneuraminic acid pH 7.0 and 37°C Bacteroides thetaiotaomicron