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Literature summary extracted from

  • Yoshigi, N.; Okada, Y.; Maeba, H.; Sahara, H.; Tamaki, T.
    Construction of a plasmid used for the expression of a sevenfold-mutant barley beta-amylase with increased thermostability in Escherichia coli and properties of the sevenfold-mutant beta-amylase (1995), J. Biochem., 118, 562-567.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.2 expression of a sevenfold mutant beta-amylase, M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.2 M185L/S295A/I297V/S350P/S351P/Q352D/A376S the mutant enzyme acquires enhanced thermostability, but its function as beta-amylase is unchanged. The mutant is stable at pH-values up to 12.5, while the original recombinant enzyme is unstable at pH-values above pH 9.5 Hordeum vulgare

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.2 Ag+ 1 mM, almost complete inhibition of mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 Cd2+ 1 mM, almost complete inhibition of barley enzyme and recombinant enzyme, less inhibitory towards mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 Cu2+ 1 mM, almost complete inhibition of mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 Hg2+ 1 mM, almost complete inhibition of mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 PCMB
-
 Glycine max
3.2.1.2 PCMB 0.1 mM, complete inhibition of mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 Zn2+ 1 mM, almost complete inhibition of barley enzyme and recombinant enzyme, less inhibitory towards mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.2 0.491
-
 amylose DP = 17, mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 1.83
-
 maltoheptaose mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 2
-
 maltohexaose mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 2.83
-
 maltopentaose mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare
3.2.1.2 4.17
-
 maltotetraose mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.2 56000
-
 x * 56000, mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S, SDS-PAGE Hordeum vulgare
3.2.1.2 59144
-
 x * 59144, mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S, calculation from amino acid sequence Hordeum vulgare

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.2 Glycine max
-
-
-
3.2.1.2 Hordeum vulgare
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.2 amylopectin + H2O
-
 Hordeum vulgare beta-maltose + ?
-
 ?
3.2.1.2 amylopectin + H2O
-
 Glycine max beta-maltose + ?
-
 ?
3.2.1.2 amylose + H2O DP = 17 Hordeum vulgare beta-maltose
-
 ?
3.2.1.2 glycogen + H2O from oyster Hordeum vulgare beta-maltose
-
 ?
3.2.1.2 maltoheptaose + H2O
-
 Hordeum vulgare ?
-
 ?
3.2.1.2 maltohexaose + H2O
-
 Hordeum vulgare ?
-
 ?
3.2.1.2 maltopentaose + H2O
-
 Hordeum vulgare ?
-
 ?
3.2.1.2 maltotetraose + H2O
-
 Hordeum vulgare ?
-
 ?
3.2.1.2 starch + H2O soluble Hordeum vulgare maltose + ?
-
 ?
3.2.1.2 starch + H2O soluble Glycine max maltose + ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.2 ?
-
 Glycine max
3.2.1.2 ? x * 59144, mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S, calculation from amino acid sequence Hordeum vulgare
3.2.1.2 ? x * 56000, mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S, SDS-PAGE Hordeum vulgare

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.2 55
-
 original and recombinant enzyme Hordeum vulgare
3.2.1.2 60
-
-
 Glycine max
3.2.1.2 65
-
 mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S Hordeum vulgare

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.2 57
-
 30 min, 50% loss of activity Hordeum vulgare
3.2.1.2 57
-
 30 min, recombinant enzyme, 50% loss of activity Hordeum vulgare
3.2.1.2 63
-
 30 min, 50% loss of activity Glycine max
3.2.1.2 69
-
 30 min, mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S, 50% inactivation Hordeum vulgare

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.2 5
-
-
 Hordeum vulgare

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.2 9.5
-
 unstable above, original recombinant enzyme Hordeum vulgare
3.2.1.2 12.5
-
 mutant enzyme M185L/S295A/I297V/S350P/S351P/Q352D/A376S is stable up to Hordeum vulgare