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Literature summary for 3.2.1.26 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
Search for more literature for 3.2.1.26

Cloned(Commentary)

Cloned (Comment) Organism
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

Protein Variants Comment Organism
C317A 2.5fold invertase activity increase, 20.7% activity with substrate raffinose compared to wild-type Bambusa oldhamii
D135E no invertase activity Bambusa oldhamii
D135N no invertase activity Bambusa oldhamii
D260E no invertase activity Bambusa oldhamii
D260N no invertase activity Bambusa oldhamii
E316A no invertase activity Bambusa oldhamii
E316Q 67.3% invertase activity retained, 15.5% activity with substrate raffinose compared to wild-type Bambusa oldhamii
R259K no invertase activity Bambusa oldhamii
Rs59L no invertase activity Bambusa oldhamii
W159F 4.7fold invertase activity increase, 41% activity with substrate raffinose compared to wild-type Bambusa oldhamii
W159L 11.7% invertase activity retained, 33.2% activity with substrate raffinose compared to wild-type Bambusa oldhamii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
53.68
-
 sucrose pH 5.0, 37°C, wild-type enzyme Bambusa oldhamii
140.7
-
 sucrose pH 5.0, 37°C, mutant enzyme C317A Bambusa oldhamii
142.6
-
 sucrose pH 5.0, 37°C, mutant enzyme E316Q Bambusa oldhamii
169.8
-
 sucrose pH 5.0, 37°C, mutant enzyme W159F Bambusa oldhamii
213.6
-
 sucrose pH 5.0, 37°C, mutant enzyme W159L Bambusa oldhamii

Localization

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Bambusa oldhamii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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
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
6.58
-
 pH 5.0, 37°C, wild-type enzyme with substrate sucrose Bambusa oldhamii
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
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
raffinose + H2O lower activity of all mutants than of wild-type Bambusa oldhamii D-fructose + melibiose
-
 ?
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

Synonyms Comment Organism
Vacuolar invertase Bobetafruct3 Bambusa oldhamii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
57.86
-
 sucrose pH 5.0, 37°C, mutant enzyme W159L Bambusa oldhamii
338.4
-
 sucrose pH 5.0, 37°C, mutant enzyme E316Q Bambusa oldhamii
501.7
-
 sucrose pH 5.0, 37°C, wild-type enzyme Bambusa oldhamii
1243
-
 sucrose pH 5.0, 37°C, mutant enzyme C317A Bambusa oldhamii
2342
-
 sucrose pH 5.0, 37°C, mutant enzyme W159F Bambusa oldhamii

pH Optimum

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