Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain AD202 | Streptococcus pneumoniae R6 |
Protein Variants | Comment | Organism |
---|---|---|
D658A | site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme | Streptococcus pneumoniae R6 |
D764A | site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme | Streptococcus pneumoniae R6 |
D764F | site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme | Streptococcus pneumoniae R6 |
E796A | site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme | Streptococcus pneumoniae R6 |
E796Q | site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme | Streptococcus pneumoniae R6 |
additional information | construction of a DELTA1-39 deletion and DELTA1-39,DELTA1568-1767 deletion mutants | Streptococcus pneumoniae R6 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analysis, overview | Streptococcus pneumoniae R6 | |
0.0012 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796Q | Streptococcus pneumoniae R6 | |
0.0013 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
0.021 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
0.033 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant D764A | Streptococcus pneumoniae R6 | |
0.034 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 | |
0.048 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39,DELTA1568-1767 deletion mutant | Streptococcus pneumoniae R6 | |
0.062 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 | |
0.212 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796Q | Streptococcus pneumoniae R6 | |
2.1 | - |
2,4-dinitrophenyl-GalNAc-alpha-R | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
2.4 | - |
2,4-dinitrophenyl-GalNAc-alpha-R | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
NaCl | effects on kinetics, overview | Streptococcus pneumoniae R6 | |
NaN3 | effects on kinetics, overview | Streptococcus pneumoniae R6 | |
Sodium formate | effects on kinetics, overview | Streptococcus pneumoniae R6 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Streptococcus pneumoniae R6 | SpGH101 specifically removes an O-linked disaccharide Gal-beta-1,3-GalNAc-alpha from glycoproteins | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus pneumoniae R6 | Q8DR60 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain AD202 by anion exchange chromatography, ultrafiltration, gel filtration, and hydrophobic interaction chromatography | Streptococcus pneumoniae R6 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc + H2O | - |
Streptococcus pneumoniae R6 | 2,4-dinitrophenol + beta-D-Gal-(1->3)-alpha-D-GalNAc | - |
? | |
2,4-dinitrophenyl-GalNAc-alpha-R + H2O | - |
Streptococcus pneumoniae R6 | 2,4-dinitrophenol + GalNAc-alpha-R | - |
? | |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc + H2O | - |
Streptococcus pneumoniae R6 | 4-nitrophenol + beta-D-Gal-(1->3)-alpha-D-GalNAc | - |
? | |
additional information | SpGH101 specifically removes an O-linked disaccharide Gal-beta-1,3-GalNAc-alpha from glycoproteins | Streptococcus pneumoniae R6 | ? | - |
? | |
additional information | the enzyme contains several putative carbohydrate binding modules, structure-function relationship, D764 and E796 are the nucleophile and general acid-base residues, respectively, overview | Streptococcus pneumoniae R6 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | analysis of the three-dimensional structure of the recombinant protein, the enzyme contains several putative carbohydrate binding modules, structure-function relationship, overview | Streptococcus pneumoniae R6 |
Synonyms | Comment | Organism |
---|---|---|
endo-alpha-N-acetylgalactosaminidase | - |
Streptococcus pneumoniae R6 |
More | the enzyme belongs to the glycoside hydrolase family 101, GH101 | Streptococcus pneumoniae R6 |
SpGH101 | - |
Streptococcus pneumoniae R6 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptococcus pneumoniae R6 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.76 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant D764A | Streptococcus pneumoniae R6 | |
1 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
1.6 | - |
2,4-dinitrophenyl-GalNAc-alpha-R | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 | |
2.1 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796Q | Streptococcus pneumoniae R6 | |
5 | - |
2,4-dinitrophenyl-GalNAc-alpha-R | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
14 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796Q | Streptococcus pneumoniae R6 | |
18 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
541 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 | |
579 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 | |
748 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39,DELTA1568-1767 deletion mutant | Streptococcus pneumoniae R6 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
bell-shaped dependence of kcat/Km upon pH | Streptococcus pneumoniae R6 |
6.5 | - |
assay at | Streptococcus pneumoniae R6 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0006 | - |
2,4-dinitrophenyl-GalNAc-alpha-R | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 | |
0.002 | - |
2,4-dinitrophenyl-GalNAc-alpha-R | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
0.0099 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796Q | Streptococcus pneumoniae R6 | |
0.023 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant D764A | Streptococcus pneumoniae R6 | |
0.048 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
0.987 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant D658A | Streptococcus pneumoniae R6 | |
9.3 | - |
4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 | |
11.7 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796Q | Streptococcus pneumoniae R6 | |
13.8 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, mutant E796A | Streptococcus pneumoniae R6 | |
15.6 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39,DELTA1568-1767 deletion mutant | Streptococcus pneumoniae R6 | |
15.9 | - |
2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc | pH 6.5, 37°C, DELTA1-39 deletion mutant | Streptococcus pneumoniae R6 |