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Literature summary extracted from

  • Juge, N.; Nohr, J.; Le Gal-Coeffet, M.F.; Kramhoft, B.; Furniss, C.S.; Planchot, V.; Archer, D.B.; Williamson, G.; Svensson, B.
    The activity of barley alpha-amylase on starch granules is enhanced by fusion of a starch binding domain from Aspergillus niger glucoamylase (2006), Biochim. Biophys. Acta, 1764, 275-284.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.1 AMY1, subcloning in Escherichia coli strain DH5alpha, expression of the alpha-amylase C-terminally fused to the Aspergillus niger glucoamlyase starch binding domain in Aspergillus niger strain AB4.1, the mutant enzyme is secreted to the culture medium due to the signal peptide of the barley alpha-amylase Hordeum vulgare

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.1 additional information construction of a mutant alpha-amylase, containing its signal peptide, which is fused to the starch binding domain, SBD, of the glucoamylase GA-I of Aspergillus niger via a 37 amino acid GA-I linker segment, the activity of the fusion protein is 2fold enhanced with amylose, and with starch at low concentration, not at high concentration, compared to the wild-type enzyme Hordeum vulgare

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.1 additional information
-
 additional information kinetics Hordeum vulgare
3.2.1.1 0.23
-
 amylose DP440 pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme Hordeum vulgare
3.2.1.1 0.37
-
 amylose DP17 pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme Hordeum vulgare
3.2.1.1 2.3
-
 2-chloro-4-nitrophenyl alpha-D-maltoheptaoside pH 6.8, 30°C, recombinant AMY1-SBD fusion enzyme Hordeum vulgare

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.1 Ca2+
-
 Hordeum vulgare

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.1 68900
-
 x * 68900, recombinant AMY1-SBD fusion enzyme, amino acid sequence calculation, x * 75000, recombinant glycosylated AMY1-SBD fusion enzyme, SDS-PAGE Hordeum vulgare
3.2.1.1 75000
-
 x * 68900, recombinant AMY1-SBD fusion enzyme, amino acid sequence calculation, x * 75000, recombinant glycosylated AMY1-SBD fusion enzyme, SDS-PAGE Hordeum vulgare

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.1 amylose + H2O Hordeum vulgare
-
 malto-oligosaccharides
-
 ?
3.2.1.1 starch + H2O Hordeum vulgare
-
 malto-oligosaccharides
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.1 Hordeum vulgare
-
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.2.1.1 glycoprotein the recombinant AMY1-SBD fusion enzyme is O-glycosylated at aminoacid residues 471-509 Hordeum vulgare

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.1 recombinant AMY1-SBD fusion enzyme from Aspergillus niger strain AB4.1 by beta-cyclodextrin affinity chromatography Hordeum vulgare

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.1 additional information
-
 activity of recombinant AMY1-SBD fusion enzyme towards different starch substrates Hordeum vulgare

Storage Stability

EC Number Storage Stability Organism
3.2.1.1 4°C, concentrated purified recombinant AMY1-SBD fusion enzyme, 50 mM MES, pH 6.5, 5 mM CaCl2, stable Hordeum vulgare

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.1 2-chloro-4-nitrophenyl alpha-D-maltoheptaoside + H2O
-
 Hordeum vulgare 2-chloro-4-nitrophenol + alpha-D-maltoheptaose
-
 ?
3.2.1.1 amylose + H2O
-
 Hordeum vulgare malto-oligosaccharides
-
 ?
3.2.1.1 amylose + H2O amylose DP440 and amylose DP17 Hordeum vulgare malto-oligosaccharides
-
 ?
3.2.1.1 amylose DP17 + H2O
-
 Hordeum vulgare ?
-
 ?
3.2.1.1 amylose DP440 + H2O
-
 Hordeum vulgare ?
-
 ?
3.2.1.1 starch + H2O
-
 Hordeum vulgare malto-oligosaccharides
-
 ?
3.2.1.1 starch + H2O starch granules, high affinity for the substrate is mediated by the enzyme's separate starch binding domain, SBD Hordeum vulgare malto-oligosaccharides
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.1 ? x * 68900, recombinant AMY1-SBD fusion enzyme, amino acid sequence calculation, x * 75000, recombinant glycosylated AMY1-SBD fusion enzyme, SDS-PAGE Hordeum vulgare

Synonyms

EC Number Synonyms Comment Organism
3.2.1.1 AMY1
-
 Hordeum vulgare

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.1 30
-
 assay at, substrate 2-chloro-4-nitrophenyl alpha-D-maltoheptoside Hordeum vulgare
3.2.1.1 37
-
 assay at, substrate starch and amylose Hordeum vulgare

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.1 132
-
 2-chloro-4-nitrophenyl alpha-D-maltoheptaoside pH 6.8, 30°C, recombinant AMY1-SBD fusion enzyme Hordeum vulgare
3.2.1.1 209
-
 amylose DP440 pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme Hordeum vulgare
3.2.1.1 225
-
 amylose DP17 pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme Hordeum vulgare

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.1 5.5
-
 assay at, substrate starch and amylose Hordeum vulgare
3.2.1.1 6.8
-
 assay at, substrate 2-chloro-4-nitrophenyl alpha-D-maltoheptoside Hordeum vulgare

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.2.1.1 Hordeum vulgare recombinant AMY1-SBD fusion enzyme, amino acid sequence calculation
-
 4.74