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Literature summary extracted from

  • Vester-Christensen, M.B.; Hachem, M.A.; Naested, H.; Svensson, B.
    Secretory expression of functional barley limit dextrinase by Pichia pastoris using high cell-density fermentation (2010), Protein Expr. Purif., 69, 112-119.
    View publication on PubMed

Application

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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.142 expression in Pichia pastoris Hordeum vulgare

KM Value [mM]

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

Molecular Weight [Da]

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

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.142 Hordeum vulgare O48541
-
-

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.142 pullulan + H2O
-
 Hordeum vulgare ?
-
 ?

Subunits

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

Turnover Number [1/s]

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

kcat/KM [mM/s]

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