EC Number | Cloned (Comment) | Organism |
---|---|---|
3.2.1.26 | Pichia pastoris X-33, His-6-tagged recombinant wild-type and mutant cDNA in plasmid pBolT3 in expression vector pPICZalphaB, 30°C, buffered glycerol-complex medium, pH 6.0 | Bambusa oldhamii |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.2.1.26 | C317A | 2.5fold invertase activity increase, 20.7% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
3.2.1.26 | D135E | no invertase activity | Bambusa oldhamii |
3.2.1.26 | D135N | no invertase activity | Bambusa oldhamii |
3.2.1.26 | D260E | no invertase activity | Bambusa oldhamii |
3.2.1.26 | D260N | no invertase activity | Bambusa oldhamii |
3.2.1.26 | E316A | no invertase activity | Bambusa oldhamii |
3.2.1.26 | E316Q | 67.3% invertase activity retained, 15.5% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
3.2.1.26 | R259K | no invertase activity | Bambusa oldhamii |
3.2.1.26 | Rs59L | no invertase activity | Bambusa oldhamii |
3.2.1.26 | W159F | 4.7fold invertase activity increase, 41% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
3.2.1.26 | W159L | 11.7% invertase activity retained, 33.2% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.26 | 53.68 | - |
sucrose | pH 5.0, 37°C, wild-type enzyme | Bambusa oldhamii | |
3.2.1.26 | 140.7 | - |
sucrose | pH 5.0, 37°C, mutant enzyme C317A | Bambusa oldhamii | |
3.2.1.26 | 142.6 | - |
sucrose | pH 5.0, 37°C, mutant enzyme E316Q | Bambusa oldhamii | |
3.2.1.26 | 169.8 | - |
sucrose | pH 5.0, 37°C, mutant enzyme W159F | Bambusa oldhamii | |
3.2.1.26 | 213.6 | - |
sucrose | pH 5.0, 37°C, mutant enzyme W159L | Bambusa oldhamii |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
3.2.1.26 | vacuole | - |
Bambusa oldhamii | 5773 | - |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.2.1.26 | 76200 | - |
gel filtration chromatography | Bambusa oldhamii |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.2.1.26 | Bambusa oldhamii | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.2.1.26 | supernatant of medium of methanol-induced cell culture precipitated by 80% saturation of ammonium sulfate, centrifuged, resuspended in sodium phosphate buffer, pH 7, dialyzed, and mixed with cobalt-based IMAC resin for 1 h at 4°C, packed into a column, eluted with imidazole | Bambusa oldhamii |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.2.1.26 | 0.768 | - |
pH 5.0, 37°C, mutant enzyme W159L with substrate sucrose, 33.2% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
3.2.1.26 | 4.43 | - |
pH 5.0, 37°C, mutant enzyme E316Q with substrate sucrose, 15.5% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
3.2.1.26 | 6.58 | - |
pH 5.0, 37°C, wild-type enzyme with substrate sucrose | Bambusa oldhamii |
3.2.1.26 | 16.3 | - |
pH 5.0, 37°C, mutant enzyme C317A with substrate sucrose, 20.7% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
3.2.1.26 | 30.73 | - |
pH 5.0, 37°C, mutant enzyme W159F with substrate sucrose, 41% activity with substrate raffinose compared to wild-type | Bambusa oldhamii |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.1.26 | raffinose + H2O | lower activity of all mutants than of wild-type | Bambusa oldhamii | D-fructose + melibiose | - |
? | |
3.2.1.26 | sucrose + H2O | 37 degrees Celsius, 0.1 M sodium acetate, pH 5, no hydrolysis of cellobiose, maltose, lactose, inulin | Bambusa oldhamii | D-fructose + D-glucose | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.2.1.26 | Vacuolar invertase | Bobetafruct3 | Bambusa oldhamii |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.26 | 57.86 | - |
sucrose | pH 5.0, 37°C, mutant enzyme W159L | Bambusa oldhamii | |
3.2.1.26 | 338.4 | - |
sucrose | pH 5.0, 37°C, mutant enzyme E316Q | Bambusa oldhamii | |
3.2.1.26 | 501.7 | - |
sucrose | pH 5.0, 37°C, wild-type enzyme | Bambusa oldhamii | |
3.2.1.26 | 1243 | - |
sucrose | pH 5.0, 37°C, mutant enzyme C317A | Bambusa oldhamii | |
3.2.1.26 | 2342 | - |
sucrose | pH 5.0, 37°C, mutant enzyme W159F | Bambusa oldhamii |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.2.1.26 | 5 | - |
wild-type and mutant enzymes | Bambusa oldhamii |