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