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Literature summary for 3.2.1.1 extracted from

  • Ghollasi, M.; Khajeh, K.; Naderi-Manesh, H.; Ghasemi, A.
    Engineering of a Bacillus alpha-amylase with improved thermostability and calcium independency (2010), Appl. Biochem. Biotechnol., 162, 444-459.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Priestia megaterium

Protein Variants

Protein Variants Comment Organism
A53S in comparison to the wild type, calcium ion has more effect on the catalytic efficiency, kcat/Km, and half-life (at 60°C) of A53S mutant, although the overall activity (kcat/Km) has not improved, about 80% of activity is maintained in the case of A53S mutant Priestia megaterium
H58I calcium-independent mutant possessing high thermostability, in the absence of CaCl2, the H58I mutant is much more stable than the wild type and there is a 7fold increase in the residual activity of H58I mutant after 30 min of incubation as compared to the wild type Priestia megaterium

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ in the presence of calcium, the affinity of the enzymes (wild type and mutants) toward starch is increased, the thermostability of the wild type and A53S mutant is calcium dependent at different temperatures, in the presence of 2 mM CaCl2 at 60°C, the percentage of residual activity in both the wild type and A53S mutant changes after 30 min of incubation Priestia megaterium

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
57000
-
predicted from amino acid sequence, SDS-PAGE Priestia megaterium

Organism

Organism UniProt Comment Textmining
Priestia megaterium P20845
-
-
Priestia megaterium WHO P20845
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography, gel filtration Priestia megaterium

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
starch + H2O
-
Priestia megaterium malto-oligosaccharides
-
?
starch + H2O
-
Priestia megaterium WHO malto-oligosaccharides
-
?

Synonyms

Synonyms Comment Organism
1,4-alpha-D-glucan glucanohydrolase
-
Priestia megaterium

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
in the absence of Ca2+ Priestia megaterium
60.7
-
in the presence of 5 mM Ca2+ Priestia megaterium

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
9.8
-
starch mutant enzyme A53S, in the presence of 5 mM Ca2+, in 20 mM Tris-HCl buffer (pH 7.2), at 30°C Priestia megaterium
19.9
-
starch wild type enzyme, in the presence of 5 mM Ca2+, in 20 mM Tris-HCl buffer (pH 7.2), at 30°C Priestia megaterium
20.7
-
starch mutant enzyme H58I, in the presence of 5 mM Ca2+, in 20 mM Tris-HCl buffer (pH 7.2), at 30°C Priestia megaterium
22.4
-
starch mutant enzyme A53S, in the absence of Ca2+, in 20 mM Tris-HCl buffer (pH 7.2), at 30°C Priestia megaterium
25.7
-
starch wild type enzyme, in the absence of Ca2+, in 20 mM Tris-HCl buffer (pH 7.2), at 30°C Priestia megaterium
27.4
-
starch mutant enzyme H58I, in the absence of Ca2+, in 20 mM Tris-HCl buffer (pH 7.2), at 30°C Priestia megaterium