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Literature summary for 3.2.1.133 extracted from

  • Sun, Y.; Duan, X.; Wang, L.; Wu, J.
    Enhanced maltose production through mutagenesis of acceptor binding subsite +2 in Bacillus stearothermophilus maltogenic amylase (2016), J. Biotechnol., 217, 53-61 .
    View publication on PubMed

Application

Application Comment Organism
food industry maltogenic amylases are used to decrease the maltotriose content of high maltose syrups. However, due to the interplay between the hydrolysis and transglycosylation activities of maltogenic amylases, the maltotriose contents of these syrups are still greater than that necessary for pure maltose preparation. Mutant enzyme W177S, shows decreased transglycosylation activity and enhanced maltose production. It will deliver performance superior to that of the wild-type under industrial conditions Geobacillus stearothermophilus

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli strains BL21(DE3) Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
W177F transglycosylation activities of the mutant enzyme decreases by 18% as the hydrophilicity of the residue at position 177 increases. The mutant enzyme exhibits notable enhancements in maltose production. The maltotriose content is substantially lower than that of the syrup produced using the wild-type enzyme Geobacillus stearothermophilus
W177L transglycosylation activities of the mutant enzyme decreases by 37% as the hydrophilicity of the residue at position 177 increases. The mutant enzyme exhibits notable enhancements in maltose production. The maltotriose content is substantially lower than that of the syrup produced using the wild-type enzyme Geobacillus stearothermophilus
W177N transglycosylation activities of the mutant enzyme decreases by 45% as the hydrophilicity of the residue at position 177 increases. The mutant enzyme exhibits notable enhancements in maltose production. The maltotriose content is substantially lower than that of the syrup produced using the wild-type enzyme Geobacillus stearothermophilus
W177S transglycosylation activities of the mutant enzyme decreases by 52% as the hydrophilicity of the residue at position 177 increases. The mutant enzyme exhibits notable enhancements in maltose production. The maltotriose content is substantially lower than that of the syrup produced using the wild-type enzyme Geobacillus stearothermophilus
W177Y transglycosylation activities of the mutant enzyme decreases by 20% as the hydrophilicity of the residue at position 177 increases. The mutant enzyme exhibits notable enhancements in maltose production. The maltotriose content is substantially lower than that of the syrup produced using the wild-type enzyme Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
12.3
-
maltotriose pH 5.5, 60°C, wild-type enzyme Geobacillus stearothermophilus
16.4
-
maltotriose pH 5.5, 60°C, mutant enzyme W177F Geobacillus stearothermophilus
22
-
maltotriose pH 5.5, 60°C, mutant enzyme W177Y Geobacillus stearothermophilus
28
-
maltotriose pH 5.5, 60°C, mutant enzyme W177L Geobacillus stearothermophilus
31.7
-
maltotriose pH 5.5, 60°C, mutant enzyme W177N Geobacillus stearothermophilus
35.3
-
maltotriose pH 5.5, 60°C, mutant enzyme W177S Geobacillus stearothermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
69000
-
SDS-PAGE Geobacillus stearothermophilus

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus P19531
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
maltotriose + H2O
-
Geobacillus stearothermophilus alpha-maltose + glucose
-
?

Subunits

Subunits Comment Organism
? x * 69000, SDS-PAGE Geobacillus stearothermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
hydrolysis of maltotriose, mutant enzyme W177Y Geobacillus stearothermophilus
60
-
hydrolysis of maltotriose, wild-type enzyme and mutant enzymes W177F, W177L, W177N, and W177S Geobacillus stearothermophilus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 70 30°C: about 40% of maximal activity, 70°C: about 65% of maximal activity, hydrolysis of maltotriose, mutant enzyme W177Y Geobacillus stearothermophilus
50 80 50°C: about 70% of maximal activity, 80°C: about 40% of maximal activity, hydrolysis of maltotriose, wild-type enzyme Geobacillus stearothermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
half-life: 325 h (wild-type enzyme), 95 h (mutant enzyme W177F), 250 h (mutant enzyme W177Y), 200 h (mutant enzyme W177L), 150 h (mutant enzyme W177N), 192 h (mutant enzyme W177S) Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
70.97
-
maltotriose pH 5.5, 60°C, mutant enzyme W177F Geobacillus stearothermophilus
108.25
-
maltotriose pH 5.5, 60°C, mutant enzyme W177Y Geobacillus stearothermophilus
126.2
-
maltotriose pH 5.5, 60°C, mutant enzyme W177L Geobacillus stearothermophilus
137.2
-
maltotriose pH 5.5, 60°C, wild-type enzyme Geobacillus stearothermophilus
150.98
-
maltotriose pH 5.5, 60°C, mutant enzyme W177N Geobacillus stearothermophilus
182.82
-
maltotriose pH 5.5, 60°C, mutant enzyme W177S Geobacillus stearothermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
hydrolysis of maltotriose, wild-type enzyme and mutant enzymes W177F, W177L, W177N, W177Y and W177S Geobacillus stearothermophilus

pH Range

pH Minimum pH Maximum Comment Organism
4 6.5 pH 4.0: about 65% of maximal activity, pH 6.5: about 60% of maximal activity, hydrolysis of maltotriose, wild-type enzyme Geobacillus stearothermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
261.3
-
maltotriose pH 5.5, 60°C, mutant enzyme W177F Geobacillus stearothermophilus
270.4
-
maltotriose pH 5.5, 60°C, mutant enzyme W177L Geobacillus stearothermophilus
285.8
-
maltotriose pH 5.5, 60°C, mutant enzyme W177N Geobacillus stearothermophilus
295.2
-
maltotriose pH 5.5, 60°C, mutant enzyme W177Y Geobacillus stearothermophilus
310.7
-
maltotriose pH 5.5, 60°C, mutant enzyme W177S Geobacillus stearothermophilus
669.2
-
maltotriose pH 5.5, 60°C, wild-type enzyme Geobacillus stearothermophilus