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Literature summary for 3.2.1.133 extracted from

  • Mehta, D.; Satyanarayana, T.
    Dimerization mediates thermo-adaptation, substrate affinity and transglycosylation in a highly thermostable maltogenic amylase of Geobacillus thermoleovorans (2013), PLoS ONE, 8, e73612.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.133

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Geobacillus thermoleovorans

Protein Variants

Protein Variants Comment Organism
D109A the mutant shows increased affinity towards amylose, amylopectin and starch, and a decreased affinity towards alpha- and beta-cyclodextrin Geobacillus thermoleovorans
D109E the mutant does not show any effect on the binding affinity and substrate hydrolytic efficiency towards alpha- and beta-cyclodextrin but a strong decline in the affinity and substrate hydrolytic efficiency of the mutant enzyme towards amylopectin Geobacillus thermoleovorans

Inhibitors

Inhibitors Comment Organism Structure
2-mercaptoethanol 69.9% residual activity at 10 mM Geobacillus thermoleovorans
Cu2+ complete inhibition at 1 mM Geobacillus thermoleovorans
diethyl dicarbonate 24.5% residual activity at 10 mM Geobacillus thermoleovorans
Hg2+ complete inhibition at 1 mM Geobacillus thermoleovorans
Mg2+ 84.21% residual activity at 1 mM Geobacillus thermoleovorans
additional information EGTA and EDTA have no observable effect on the activity Geobacillus thermoleovorans
N-bromosuccinamide complete inhibition at 1 mM Geobacillus thermoleovorans
Woodward's reagent K 72.4% residual activity at 10 mM Geobacillus thermoleovorans

Metals/Ions

Metals/Ions Comment Organism Structure
Al3+ 117.4% activity at 1 mM Geobacillus thermoleovorans
Co2+ 131.2% activity at 1 mM Geobacillus thermoleovorans
Mn2+ 145.1% activity at 1 mM Geobacillus thermoleovorans
additional information K+, Ni2+, Ca2+, and Ba2+ do not exert any observable effect on enzyme activity Geobacillus thermoleovorans
Na+ 123% activity at 1 mM Geobacillus thermoleovorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
72500
-
 2 * 72500, SDS-PAGE Geobacillus thermoleovorans
145000
-
 dimer, gel filtration Geobacillus thermoleovorans
725000
-
 1 * 725000, SDS_PAGE Geobacillus thermoleovorans

Organism

Organism UniProt Comment Textmining
Geobacillus thermoleovorans G8N704
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-NTA agarose resin column chromatography, and Sephacryl S-200 gel filtration Geobacillus thermoleovorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acarbose + H2O
-
 Geobacillus thermoleovorans D-glucose + acarviosine-glucose
-
 ?
alpha-cyclodextrin + H2O
-
 Geobacillus thermoleovorans maltose + D-glucose
-
 ?
amylopectin + H2O
-
 Geobacillus thermoleovorans maltose + ?
-
 ?
amylose + H2O
-
 Geobacillus thermoleovorans maltose + ?
-
 ?
beta-cyclodextrin + H2O
-
 Geobacillus thermoleovorans maltose + D-glucose
-
 ?
additional information the dimeric enzyme transglycosylates hydrolytic products of G4/G5 and acarbose, while the monomeric form does not because of the lack of extra sugar-binding space formed due to dimerization Geobacillus thermoleovorans ?
-
 ?
pullulan + H2O
-
 Geobacillus thermoleovorans ?
-
 ?
soluble starch + H2O the enzyme displays less hydrolytic action on raw starches than on soluble starch Geobacillus thermoleovorans maltose + ?
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 72500, SDS-PAGE Geobacillus thermoleovorans
monomer 1 * 725000, SDS_PAGE Geobacillus thermoleovorans

Synonyms

Synonyms Comment Organism
glucan-1,4-alpha-maltohydrolase
-
 Geobacillus thermoleovorans
maltogenic amylase
-
 Geobacillus thermoleovorans
MAmy
-
 Geobacillus thermoleovorans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
 Geobacillus thermoleovorans

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 100 about 35% activity at 40°C, about 60% activity at 50°C, about 80% activity at 90°C, about 35% activity at 70°C, 100% activity at 80°C, about 97% activity at 90°C, and about 75% activity at 100°C Geobacillus thermoleovorans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
90
-
 the half-life values of the monomeric enzyme at 80, 90 and 100°C are 35.7, 8.2 and 3.12 h, respectively. The half-life values of the dimeric enzyme at 80, 90 and 100°C are 55.7, 12.63 and 3.58 h, respectively Geobacillus thermoleovorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.005
-
 amylopectin mutant enzyme D109E, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.021
-
 soluble starch mutant enzyme D109E, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.022
-
 amylose mutant enzyme D109E, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.04
-
 amylose dimeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.04
-
 amylopectin dimeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.04
-
 soluble starch dimeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.1
-
 alpha-cyclodextrin monomeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.1
-
 amylopectin mutant enzyme D109A, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.12
-
 beta-cyclodextrin monomeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.12
-
 soluble starch mutant enzyme D109A, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.13
-
 amylose mutant enzyme D109A, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.17
-
 amylopectin monomeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.2
-
 soluble starch monomeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.21
-
 amylose monomeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
0.49
-
 beta-cyclodextrin mutant enzyme D109A at pH 7.0 and 80°C Geobacillus thermoleovorans
0.66
-
 beta-cyclodextrin mutant enzyme D109E at pH 7.0 and 80°C Geobacillus thermoleovorans
0.75
-
 alpha-cyclodextrin mutant enzyme D109A, at pH 7.0 and 80°C Geobacillus thermoleovorans
1.33
-
 beta-cyclodextrin dimeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans
1.39
-
 alpha-cyclodextrin mutant enzyme D109E, at pH 7.0 and 80°C Geobacillus thermoleovorans
2.78
-
 alpha-cyclodextrin dimeric enzyme form, at pH 7.0 and 80°C Geobacillus thermoleovorans

pH Range

pH Minimum pH Maximum Comment Organism
5 9 the enzyme is active over a wide range of pH between 5.0 and 9.0 (more than 90% activity) but the activity drastically declines at pH 4.0 and 10.0 to 20 and less than 5%, respectively Geobacillus thermoleovorans

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 8 there is no loss in the activity of the enzyme till 14 days of incubation at pH 7.0 and 8.0, but it decreases drastically after 8 days at pH 5.0 and 6.0 Geobacillus thermoleovorans