Protein Variants | Comment | Organism |
---|---|---|
H98A | site-directed mutagenesis, the mutant shows affected kinetics and reduced activity compared to the wild-type enzyme | Thermobacillus xylanilyticus |
H98A/W99A | site-directed mutagenesis, the mutant shows affected kinetics and reduced activity compared to the wild-type enzyme | Thermobacillus xylanilyticus |
H98F | site-directed mutagenesis, the mutant shows affected kinetics and reduced activity compared to the wild-type enzyme | Thermobacillus xylanilyticus |
H98F/W99F | site-directed mutagenesis, the mutant shows affected kinetics and reduced activity compared to the wild-type enzyme | Thermobacillus xylanilyticus |
W99A | site-directed mutagenesis, the mutation causes the loss of a hydrogen bond and leads to an alternative binding mode that is detrimental for catalysis, altered binding of the aglycon motif in the active site, the mutant shows affected kinetics and reduced activity compared to the wild-type enzyme | Thermobacillus xylanilyticus |
W99F | site-directed mutagenesis, the mutant shows affected kinetics, reduced activity, and altered binding of the aglycon motif in the active site compared to the wild-type enzyme | Thermobacillus xylanilyticus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.72 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | wild-type enzyme, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
0.87 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
1.5 | 2 | 4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
3.4 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98F/W99F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
4.2 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant W99F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
9.1 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant W99A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
15 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98A/W99A, pH 5.8, 60°C | Thermobacillus xylanilyticus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Thermobacillus xylanilyticus | alpha-L-arabinofuranosidases are exo-acting glycoside hydrolases that catalyse the hydrolysis of terminal non-reducing alpha-L-arabinofuranosyl moieties in oligosaccharides and polysaccharides through the cleavage of 1->2 and/or 1->3 glycosidic bonds | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermobacillus xylanilyticus | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl alpha-L-arabinofuranoside + H2O | the substrate binding process is driven by favourable entropy, which is linked to the movement of the beta2alpha2 loop. Loop closure relocates the solvent-exposed W99 into a buried location, allowing its involvement in substrate binding and in the formation of a functional active site. Residue H98 has a role in the dynamic formation as catalytically operational active site, which may be a specific feature of a subset of GH51 arabinofuranosidases | Thermobacillus xylanilyticus | 4-nitrophenol + L-arabinofuranose | - |
? | |
additional information | alpha-L-arabinofuranosidases are exo-acting glycoside hydrolases that catalyse the hydrolysis of terminal non-reducing alpha-L-arabinofuranosyl moieties in oligosaccharides and polysaccharides through the cleavage of 1->2 and/or 1->3 glycosidic bonds | Thermobacillus xylanilyticus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure of the enzyme, overview | Thermobacillus xylanilyticus |
Synonyms | Comment | Organism |
---|---|---|
ABF | - |
Thermobacillus xylanilyticus |
alpha-L-arabinofuranosidase | - |
Thermobacillus xylanilyticus |
GH51 arabinofuranosidase | - |
Thermobacillus xylanilyticus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Thermobacillus xylanilyticus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
77 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant W99F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
97 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98F/W99F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
137 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant W99A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
154 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98A/W99A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
192 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
199 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
575 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | wild-type enzyme, pH 5.8, 60°C | Thermobacillus xylanilyticus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.8 | - |
assay at | Thermobacillus xylanilyticus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the family 51 glycoside hydrolase, GH51 | Thermobacillus xylanilyticus |
additional information | functional role of the mobile beta2alpha2 loop, especially of loop residues H98 and W99, analysis by site-directed mutagenesis, isothermal titration calorimetry, saturation transfer difference NMR spectroscopy, and molecular dynamics simulation, three-dimensional structure of the enzyme and its active site, overview | Thermobacillus xylanilyticus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
9.8 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98A/W99A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
15.1 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant W99A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
18 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant W99F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
28.2 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98F/W99F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
126 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98A, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
228 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | mutant H98F, pH 5.8, 60°C | Thermobacillus xylanilyticus | |
795 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | wild-type enzyme, pH 5.8, 60°C | Thermobacillus xylanilyticus |