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Literature summary for 3.2.1.1 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
Search for more literature for 3.2.1.1

Cloned(Commentary)

Cloned (Comment) Organism
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

Protein Variants Comment Organism
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]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+
-
 Hordeum vulgare

Molecular Weight [Da]

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

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

Organism

Organism UniProt Comment Textmining
Hordeum vulgare
-
-
-

Posttranslational Modification

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

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Storage Stability

Storage Stability Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-chloro-4-nitrophenyl alpha-D-maltoheptaoside + H2O
-
 Hordeum vulgare 2-chloro-4-nitrophenol + alpha-D-maltoheptaose
-
 ?
amylose + H2O
-
 Hordeum vulgare malto-oligosaccharides
-
 ?
amylose + H2O amylose DP440 and amylose DP17 Hordeum vulgare malto-oligosaccharides
-
 ?
amylose DP17 + H2O
-
 Hordeum vulgare ?
-
 ?
amylose DP440 + H2O
-
 Hordeum vulgare ?
-
 ?
starch + H2O
-
 Hordeum vulgare malto-oligosaccharides
-
 ?
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

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

Synonyms

Synonyms Comment Organism
AMY1
-
 Hordeum vulgare

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

pI Value

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