Activating Compound | Comment | Organism | Structure |
---|---|---|---|
dithiothreitol | 32% increase of activity by 10 mM dithiothreitol | Streptococcus mitis |
Cloned (Comment) | Organism |
---|---|
expressed as His-tagged enzyme in Escherichia coli MC4100 cells | Streptococcus mitis |
General Stability | Organism |
---|---|
the enzyme remains highly stable in solution or immobilized at room temperature in the absence of protein stabilizers, the His tag does not affect activity | Streptococcus mitis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | 46% residual activity at 5 mM | Streptococcus mitis | |
Fe2+ | 68% residual activity at 5 mM | Streptococcus mitis | |
additional information | enzyme activity is not inhibited by D-glucose, even at 200 mM | Streptococcus mitis | |
SDS | 1% (w/v) SDS strongly inhibits the ONPGase activity | Streptococcus mitis | |
Zn2+ | 60% residual activity at 5 mM | Streptococcus mitis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cell wall | - |
Streptococcus mitis | 5618 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
252000 | - |
analytical ultracentrifugation | Streptococcus mitis |
264000 | - |
2 * 264000, full-length enzyme, estimated from SDS-PAGE | Streptococcus mitis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus mitis | - |
- |
- |
Streptococcus mitis NCTC 12261T | - |
- |
- |
Purification (Comment) | Organism |
---|---|
the mature enzyme is purified by DEAE-cellulose column chromatography and Sephacryl S-300 gel filtration, the His-tagged enzyme is purified by Bio-Scale Mini Profinity immobilized metal affinity chromatography | Streptococcus mitis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
898 | - |
mature enzyme from crude extract, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol | Streptococcus mitis |
1093 | - |
His-tagged enzyme from crude extract, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol | Streptococcus mitis |
2444 | - |
mature enzyme after 2.7fold purification, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol | Streptococcus mitis |
2969 | - |
His-tagged enzyme after 2.7fold purification, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol | Streptococcus mitis |
Storage Stability | Organism |
---|---|
-20°C, 20 mM phosphate buffer, pH 6.9 without the addition of protein stabilizers, at least 2 years, no loss of activity | Streptococcus mitis |
25°C, 20 mM phosphate buffer, pH 6.9 without the addition of protein stabilizers, 55 days, no loss of activity | Streptococcus mitis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-nitrophenyl beta-D-galactopyranoside + H2O | about 83% activity compared to 4-nitrophenyl-beta-D-galactopyranoside | Streptococcus mitis | 2-nitrophenol + beta-D-galactose | - |
? | |
2-nitrophenyl beta-D-galactopyranoside + H2O | about 83% activity compared to 4-nitrophenyl-beta-D-galactopyranoside | Streptococcus mitis NCTC 12261T | 2-nitrophenol + beta-D-galactose | - |
? | |
4-nitrophenyl beta-D-galactopyranoside + H2O | 100% activity | Streptococcus mitis | 4-nitrophenol + beta-D-galactose | - |
? | |
4-nitrophenyl beta-D-galactopyranoside + H2O | 100% activity | Streptococcus mitis NCTC 12261T | 4-nitrophenol + beta-D-galactose | - |
? | |
lactose + H2O | about 36% activity compared to 4-nitrophenyl-beta-D-galactopyranoside | Streptococcus mitis | D-galactose + D-glucose | - |
? | |
lactose + H2O | about 36% activity compared to 4-nitrophenyl-beta-D-galactopyranoside | Streptococcus mitis NCTC 12261T | D-galactose + D-glucose | - |
? | |
lactulose + H2O | about 0.7% activity compared to 4-nitrophenyl-beta-D-galactopyranoside | Streptococcus mitis | ? | - |
? | |
lactulose + H2O | about 0.7% activity compared to 4-nitrophenyl-beta-D-galactopyranoside | Streptococcus mitis NCTC 12261T | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 2 * 264000, full-length enzyme, estimated from SDS-PAGE | Streptococcus mitis |
Synonyms | Comment | Organism |
---|---|---|
beta-D-galactoside galactohydrolase | - |
Streptococcus mitis |
ONPGase | - |
Streptococcus mitis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 40 | - |
Streptococcus mitis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
above 50°C the activity rapidly decreases | Streptococcus mitis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 6.5 | - |
Streptococcus mitis |