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Literature summary extracted from

  • Chen, T.H.; Huang, Y.C.; Yang, C.S.; Yang, C.C.; Wang, A.Y.; Sung, H.Y.
    Insights into the catalytic properties of bamboo vacuolar invertase through mutational analysis of active site residues (2009), Phytochemistry, 70, 25-31.
    View publication on PubMed

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

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

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.26 vacuole
-
 Bambusa oldhamii 5773
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.26 76200
-
 gel filtration chromatography Bambusa oldhamii

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.26 Bambusa oldhamii
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.26 Vacuolar invertase Bobetafruct3 Bambusa oldhamii

Turnover Number [1/s]

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

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.26 5
-
 wild-type and mutant enzymes Bambusa oldhamii