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Literature summary for 3.2.1.52 extracted from

  • Jiang, Y.L.; Yu, W.L.; Zhang, J.W.; Frolet, C.; Di Guilmi, A.M.; Zhou, C.Z.; Vernet, T.; Chen, Y.
    Structural basis for the substrate specificity of a novel {beta}-N-acetyl-hexosaminidase StrH from Streptococcus pneumoniae R6 (2011), J. Biol. Chem., 286, 43004-43012.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.52

Crystallization (Commentary)

Crystallization (Comment) Organism
2.1 A crystal structure of the N-terminal domain of StrH, residues Glu175-Lys642, complexed with N-acetylglucosamine. It adopts an overall structure similar to other GH20 enzymes, a (beta/alpha)8 TIM-barrel with the active site residing at the center of the beta-barrel convex side Streptococcus pneumoniae

Protein Variants

Protein Variants Comment Organism
additional information enzyme consists of a tandem repeat of two 53% sequence-identical catalytic domains designated as GH20-1 and GH20-2, respectively. Separate expression of the two domains, GH20-1 has an enzymatic activity of one-fourth compared to GH20-2 Streptococcus pneumoniae
W443A mutant of catalytic domain GH20-1, comparable Km but much lower kcat values than wild-type, enzymatic activities kcat/Km of about 1% towards 4-nitrophenyl-N-acetylglucosamine and one-fourth towards N-acetylglucosaminyl-beta-1,2-Man Streptococcus pneumoniae
W443A/W876A significant decrease in activity Streptococcus pneumoniae
W443A/W876A/Y482A/Y914A no detectable activity Streptococcus pneumoniae
W443A/Y482A mutant of catalytic domain GH20-1, no detectable activity Streptococcus pneumoniae
W443F mutant of catalytic domain GH20-1, one-sixth of wild-type activity towards 4-nitrophenyl-N-acetylglucosamine Streptococcus pneumoniae
W876A mutant of catalytic domain GH20-2, one-third of wild-type activity towards 4-nitrophenyl-N-acetylglucosamine Streptococcus pneumoniae
W876A/Y914A mutant of catalytic domain GH20-2, no detectable activity Streptococcus pneumoniae
W876F mutant of catalytic domain GH20-2, one-eighth of wild-type activity towards 4-nitrophenyl-N-acetylglucosamine Streptococcus pneumoniae
Y482A mutant of catalytic domain GH20-1, 2fold increase in Km and one-third of kcat value, leading to a much lower activity towards 4-nitrophenyl-N-acetylglucosamine than wild-type. Mutation completely abolishes the enzymatic activity towards N-acetylglucosamine-beta-1,2-Man Streptococcus pneumoniae
Y482A mutant of catalytic domain GH20-1, one-sixth of wild-type activity towards 4-nitrophenyl-N-acetylglucosamine, no activity with N-acetylglucosaminyl-beta-1,2-Man Streptococcus pneumoniae
Y482A/Y914A complete loss of activity Streptococcus pneumoniae
Y482F mutant of catalytic domain GH20-1, one-third of wild-type activity towards 4-nitrophenyl-N-acetylglucosamine Streptococcus pneumoniae
Y914A mutant of catalytic domain GH20-2, increase in kcat value Streptococcus pneumoniae
Y914F mutant of catalytic domain GH20-2, activity comparable to wild-type Streptococcus pneumoniae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0857
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443A, pH 7.5, 37°C Streptococcus pneumoniae
0.5
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering GH20-1, the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.5
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.65
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant C469Y, pH 7.5, 37°C Streptococcus pneumoniae
0.7
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering GH20-1, and the linker, pH 7.5, 37°C Streptococcus pneumoniae
0.787
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.837
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide catalytic domain GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.85
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide catalytic domain GH20-1, pH 7.5, 37°C Streptococcus pneumoniae
0.9
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant E361A, pH 7.5, 37°C Streptococcus pneumoniae
0.9
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443A/W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.95
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y914F, pH 7.5, 37°C Streptococcus pneumoniae
0.98
-
 N-acetylglucosaminyl-beta-1,2-Man construct of StrH covering GH20-1, the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
1
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W876A, pH 7.5, 37°C Streptococcus pneumoniae
1.03
-
 N-acetylglucosaminyl-beta-1,2-Man catalytic domain GH20-1, pH 7.5, 37°C Streptococcus pneumoniae
1.12
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443F, pH 7.5, 37°C Streptococcus pneumoniae
1.29
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W876F, pH 7.5, 37°C Streptococcus pneumoniae
1.3
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W443A/W876A, pH 7.5, 37°C Streptococcus pneumoniae
1.31
-
 N-acetylglucosaminyl-beta-1,2-Man catalytic domain GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
1.34
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W443A, pH 7.5, 37°C Streptococcus pneumoniae
1.5
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y482F, pH 7.5, 37°C Streptococcus pneumoniae
1.51
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W876A, pH 7.5, 37°C Streptococcus pneumoniae
1.56
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y914A, pH 7.5, 37°C Streptococcus pneumoniae
1.8
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y482A, pH 7.5, 37°C Streptococcus pneumoniae

Organism

Organism UniProt Comment Textmining
Streptococcus pneumoniae Q8DRL6 member of glycosyl hydrolase family 20
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl N-acetyl-beta-D-glucosaminide + H2O
-
 Streptococcus pneumoniae N-acetyl-beta-D-glucosamine + 4-nitrophenol
-
 ?
N-acetylglucosaminyl-beta-1,2-Man + H2O
-
 Streptococcus pneumoniae N-acetyl-beta-D-glucosamine + D-mannose
-
 ?

Synonyms

Synonyms Comment Organism
StrH
-
 Streptococcus pneumoniae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0087
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W443A, pH 7.5, 37°C Streptococcus pneumoniae
0.009
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.01
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W443A/W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.015
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443A, pH 7.5, 37°C Streptococcus pneumoniae
0.025
-
 N-acetylglucosaminyl-beta-1,2-Man catalytic domain GH20-1, pH 7.5, 37°C Streptococcus pneumoniae
0.033
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y914A, pH 7.5, 37°C Streptococcus pneumoniae
0.04
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443A/W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.1
-
 N-acetylglucosaminyl-beta-1,2-Man catalytic domain GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.11
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443F, pH 7.5, 37°C Streptococcus pneumoniae
0.4
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant E361A, pH 7.5, 37°C Streptococcus pneumoniae
0.46
-
 N-acetylglucosaminyl-beta-1,2-Man construct of StrH covering GH20-1, the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.46
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y482A, pH 7.5, 37°C Streptococcus pneumoniae
0.51
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.66
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y482F, pH 7.5, 37°C Streptococcus pneumoniae
0.98
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W876F, pH 7.5, 37°C Streptococcus pneumoniae
1.3
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide catalytic domain GH20-1, pH 7.5, 37°C Streptococcus pneumoniae
1.4
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering GH20-1, and the linker, pH 7.5, 37°C Streptococcus pneumoniae
4.7
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y914F, pH 7.5, 37°C Streptococcus pneumoniae
4.8
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide catalytic domain GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
4.9
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
5.1
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant C469Y, pH 7.5, 37°C Streptococcus pneumoniae
11
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering GH20-1, the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.006
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.0065
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W443A, pH 7.5, 37°C Streptococcus pneumoniae
0.007
-
 N-acetylglucosaminyl-beta-1,2-Man mutant W443A/W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.018
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443A, pH 7.5, 37°C Streptococcus pneumoniae
0.022
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y914A, pH 7.5, 37°C Streptococcus pneumoniae
0.024
-
 N-acetylglucosaminyl-beta-1,2-Man catalytic domain GH20-1, pH 7.5, 37°C Streptococcus pneumoniae
0.044
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443A/W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.078
-
 N-acetylglucosaminyl-beta-1,2-Man catalytic domain GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.09
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W443F, pH 7.5, 37°C Streptococcus pneumoniae
0.25
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y482A, pH 7.5, 37°C Streptococcus pneumoniae
0.4
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant E361A, pH 7.5, 37°C Streptococcus pneumoniae
0.44
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y482F, pH 7.5, 37°C Streptococcus pneumoniae
0.51
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W876A, pH 7.5, 37°C Streptococcus pneumoniae
0.645
-
 N-acetylglucosaminyl-beta-1,2-Man construct of StrH covering GH20-1, the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
0.75
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant W876F, pH 7.5, 37°C Streptococcus pneumoniae
1.5
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide catalytic domain GH20-1, pH 7.5, 37°C Streptococcus pneumoniae
2
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering GH20-1, and the linker, pH 7.5, 37°C Streptococcus pneumoniae
5
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant Y914F, pH 7.5, 37°C Streptococcus pneumoniae
5.8
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide catalytic domain GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
6.3
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae
7.8
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide mutant C469Y, pH 7.5, 37°C Streptococcus pneumoniae
22
-
 4-nitrophenyl N-acetyl-beta-D-glucosaminide construct of StrH covering GH20-1, the linker and GH20-2, pH 7.5, 37°C Streptococcus pneumoniae