EC Number | Application | Comment | Organism |
---|---|---|---|
3.2.1.142 | synthesis | expression of the limit dextrinase encoding gene fragment without signal peptide, with the Saccharomyces cerevisiae alpha-factor secretion signal under control of the alcohol oxidase 1 promoter, in Pichia pastoris leads to highly active barley limit dextrinase secreted during high cell-density fermentation. Optimization of a fedbatch fermentation procedure enables efficient production in a 5-l bioreactor, yielding 34 mg homogenous enzyme with 84% recovery | Hordeum vulgare |
EC Number | Cloned (Comment) | Organism |
---|---|---|
3.2.1.142 | expression in Pichia pastoris | Hordeum vulgare |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.142 | additional information | - |
additional information | classical Michaelis-Menten kinetics results in Km of 0.081 mg/ml, fitting the uncompetitive substrate inhibition Michaelis-Menten equation results in Km 0.16 mg/ml, pH 5.5, 40°C | Hordeum vulgare |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.2.1.142 | 97419 | - |
x * 98000, SDS-PAGE, recombinant enzyme, x * 97419, calculated | Hordeum vulgare |
3.2.1.142 | 98000 | - |
x * 98000, SDS-PAGE, recombinant enzyme, x * 97419, calculated | Hordeum vulgare |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.2.1.142 | Hordeum vulgare | O48541 | - |
- |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.2.1.142 | 14.2 | - |
pH 5.5, 40°C | Hordeum vulgare |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.1.142 | pullulan + H2O | - |
Hordeum vulgare | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.2.1.142 | ? | x * 98000, SDS-PAGE, recombinant enzyme, x * 97419, calculated | Hordeum vulgare |
3.2.1.142 | More | binding of alpha-, beta-, and gamma-cyclodextrin binding to limit dextrinase with Kd of 27.2, 0.70, and 34.7 microM, respectively | Hordeum vulgare |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.142 | 61 | - |
pullulan | classical Michaelis-Menten kinetics, pH 5.5, 40°C | Hordeum vulgare | |
3.2.1.142 | 78 | - |
pullulan | fitting the uncompetitive substrate inhibition Michaelis-Menten equation, pH 5.5, 40°C | Hordeum vulgare |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.142 | additional information | - |
additional information | classical Michaelis-Menten kinetics results in kcat/Km 753 ml/mg s, fitting the uncompetitive substrate inhibition Michaelis-Menten equation results in kcat/Km 488 ml/mg s, pH 5.5, 40°C | Hordeum vulgare |