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Literature summary for 3.2.1.52 extracted from

  • Manuel, S.G.; Ragunath, C.; Sait, H.B.; Izano, E.A.; Kaplan, J.B.; Ramasubbu, N.
    Role of active-site residues of dispersin B, a biofilm-releasing beta-hexosaminidase from a periodontal pathogen, in substrate hydrolysis (2007), FEBS J., 274, 5987-5999.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
D183N no activity with 4-methylumbelliferyl N-acetyl-beta-D-glucosamine. Kcat/KM for 4-nitrophenyl N-acetyl-beta-D-glucosamine is 13333fold lower than wild-type value. Biofilm-detachment activity is very low Aggregatibacter actinomycetemcomitans
E184Q no activity with 4-methylumbelliferyl N-acetyl-beta-D-glucosamine. Kcat/KM for 4-nitrophenyl N-acetyl-beta-D-glucosamine is 70.6fold lower than wild-type value. pH-optimum is shifted from pH 5.8 for wild-type enzyme to pH 6.8 for the mutant enzyme Aggregatibacter actinomycetemcomitans
E332Q no activity with 4-methylumbelliferyl N-acetyl-beta-D-glucosamine. Kcat/KM for 4-nitrophenyl N-acetyl-beta-D-glucosamine is 2000fold lower than wild-type value. Biofilm-detachment activity is 28fold less efficient than the wild-type Aggregatibacter actinomycetemcomitans
R27A no activity with 4-methylumbelliferyl N-acetyl-beta-D-glucosamine. Kcat/KM for 4-nitrophenyl N-acetyl-beta-D-glucosamine is 2400fold lower than wild-type value. Biofilm-detachment activity is 20fold less efficient than the wild-type Aggregatibacter actinomycetemcomitans
R27K no activity with 4-methylumbelliferyl N-acetyl-beta-D-glucosamine. Kcat/KM for 4-nitrophenyl N-acetyl-beta-D-glucosamine is 1714fold lower than wild-type value. Biofilm-detachment activity is 26fold less efficient than the wild-type Aggregatibacter actinomycetemcomitans
W237A no activity with 4-methylumbelliferyl N-acetyl-beta-D-glucosamine and 4-nitrophenyl N-acetyl-beta-D-glucosamine. Biofilm-detachment activity is 167fold less efficient than the wild-type Aggregatibacter actinomycetemcomitans
W330Y no activity with 4-methylumbelliferyl N-acetyl-beta-D-glucosamine and 4-nitrophenyl N-acetyl-beta-D-glucosamine. Biofilm-detachment activity is 19.2fold less efficient than the wild-type Aggregatibacter actinomycetemcomitans
Y187A Kcat/KM for 4-nitrophenyl N-acetyl-beta-D-glucosamine is 40fold lower than wild-type value. Kcat/KM for 4-methylumbelliferyl N-acetyl-beta-D-glucosamine is 4.9fold lower than wild-type value. Biofilm-detachment activity is 8.3fold less efficient than the wild-type Aggregatibacter actinomycetemcomitans
Y278A Kcat/KM for 4-nitrophenyl N-acetyl-beta-D-glucosamine is 923fold lower than wild-type value. Kcat/KM for 4-methylumbelliferyl N-acetyl-beta-D-glucosamine is 45fold lower than wild-type value. Biofilm-detachment activity is 11fold less efficient than the wild-type Aggregatibacter actinomycetemcomitans

Inhibitors

Inhibitors Comment Organism Structure
NAG-thiazoline mechanism-based inhibitor Aggregatibacter actinomycetemcomitans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.39
-
4-methylumbelliferyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, wild-type enzyme Aggregatibacter actinomycetemcomitans
0.59
-
4-methylumbelliferyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y187A Aggregatibacter actinomycetemcomitans
1.13
-
4-methylumbelliferyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y278A Aggregatibacter actinomycetemcomitans
3.7
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme E184Q Aggregatibacter actinomycetemcomitans
6.5
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme E332Q Aggregatibacter actinomycetemcomitans
14.5
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme R27A Aggregatibacter actinomycetemcomitans
19.4
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme D183N Aggregatibacter actinomycetemcomitans
35.9
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y278A Aggregatibacter actinomycetemcomitans
46.2
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, wild-type enzyme Aggregatibacter actinomycetemcomitans
76.2
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme R27K Aggregatibacter actinomycetemcomitans
101.1
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y187A Aggregatibacter actinomycetemcomitans

Organism

Organism UniProt Comment Textmining
Aggregatibacter actinomycetemcomitans
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Aggregatibacter actinomycetemcomitans

Storage Stability

Storage Stability Organism
-20°C, buffer comprising 50 mM sodium phosphate, pH 5.8, 50 mm NaCl and 50% glycerol, stable for several months Aggregatibacter actinomycetemcomitans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl N-acetyl-beta-D-glucosamine + H2O
-
Aggregatibacter actinomycetemcomitans 4-methylumbelliferone + N-acetyl-beta-D-glucosamine
-
?
4-nitrophenyl N-acetyl-beta-D-glucosamine + H2O
-
Aggregatibacter actinomycetemcomitans 4-nitrophenol + N-acetyl-beta-D-glucosamine
-
?

Synonyms

Synonyms Comment Organism
dispersin B
-
Aggregatibacter actinomycetemcomitans
DspB
-
Aggregatibacter actinomycetemcomitans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.000083
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme E332Q Aggregatibacter actinomycetemcomitans
0.00033
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme D183N Aggregatibacter actinomycetemcomitans
0.00038
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme R27A Aggregatibacter actinomycetemcomitans
0.00083
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme R27K Aggregatibacter actinomycetemcomitans
0.00122
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme E184Q Aggregatibacter actinomycetemcomitans
0.0015
-
4-methylumbelliferyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y278A Aggregatibacter actinomycetemcomitans
0.00417
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y278A Aggregatibacter actinomycetemcomitans
0.00717
-
4-methylumbelliferyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y187A Aggregatibacter actinomycetemcomitans
0.0235
-
4-methylumbelliferyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, wild-type enzyme Aggregatibacter actinomycetemcomitans
0.06
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, mutant enzyme Y187A Aggregatibacter actinomycetemcomitans
0.927
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, wild-type enzyme Aggregatibacter actinomycetemcomitans
6.08
-
4-nitrophenyl N-acetyl-beta-D-glucosamine 37°C, pH 5.8, wild-type enzyme Aggregatibacter actinomycetemcomitans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.8
-
wild-type enzyme Aggregatibacter actinomycetemcomitans
6.8
-
mutant enzyme E184Q Aggregatibacter actinomycetemcomitans

pH Range

pH Minimum pH Maximum Comment Organism
5 7 pH 5: about 65% of maximal activity, pH 7: about 45% of maximal activity, wild-type enzyme Aggregatibacter actinomycetemcomitans

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.147
-
-
Aggregatibacter actinomycetemcomitans NAG-thiazoline