BRENDA - Enzyme Database

Enhancement of the alcoholytic activity of alpha-amylase AmyA from Thermotoga maritima MSB8 (DSM 3109) by site-directed mutagenesis

Damian-Almazo, J.Y.; Moreno, A.; Lopez-Munguia, A.; Soberon, X.; Gonzalez-Munoz, F.; Saab-Rincon, G.; Appl. Environ. Microbiol. 74, 5168-5177 (2008)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
3.2.1.1
industry
thermal stability of the wild-type and mutant enzymes fulfill the stability requirements for industrial processes
Thermotoga maritima MSB8
Engineering
EC Number
Protein Variants
Commentary
Organism
3.2.1.1
F179V
yield a similar product profile to that of the wild-type enzyme
Thermotoga maritima MSB8
3.2.1.1
H222D
show different product spectrum than wild-type, in addition small amounts of maltotriose are produced. When methanol and butanol are used as nucleophil instead of H2O, the mutant produces more methylglucoside and butylglucoside than wild-type, respectively
Thermotoga maritima MSB8
3.2.1.1
H222E
show different product spectrum than wild-type, in addition small amounts of maltotriose are produced. When methanol is used as nucleophil instead of H2O, more methylglucoside is produced than with wild-type enzyme. Butanol produces almost equal amount in both wild-type and H222E mutant
Thermotoga maritima MSB8
3.2.1.1
H222Q
show different product spectrum than wild-type, in addition small amounts of maltotriose are produced. When methanol and butanol are used as nucleophil instead of H2O, the mutant produces more methylglucoside and butylglucoside than wild-type, respectively
Thermotoga maritima MSB8
3.2.1.1
V259W
in addition to alpha-D-glucose and maltose the product profile is broadened with small amounts of maltotriose
Thermotoga maritima MSB8
3.2.1.1
W177V
yield a similar product profile to that of the wild-type enzyme
Thermotoga maritima MSB8
3.2.1.1
Y178V
yield a similar product profile to that of the wild-type enzyme
Thermotoga maritima MSB8
Organism
EC Number
Organism
UniProt
Commentary
Textmining
3.2.1.1
Thermotoga maritima MSB8
P96107
DSM 3109, hyperthermophilic bacterium
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
3.2.1.1
wild-type and mutants H222D, H222E, H222Q, W177V, Y178V, and F179V
Thermotoga maritima MSB8
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.2.1.1
additional information
-
1000 U/mg for the wild-type enzyme and the mutants W177V, Y178V, and F179V
Thermotoga maritima MSB8
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
3.2.1.1
starch + butanol
-
690543
Thermotoga maritima MSB8
butylglucoside + alpha-D-glucose
butylglucoside: wild-type ca. 4.5 mg/ml, mutant H222Q ca. 6.5 mg/ml, mutant H222E ca. 3.5 mg/ml, mutant H222D ca. 5.5 mg/mk, identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
3.2.1.1
starch + H2O
pH 7.0, 85°C
690543
Thermotoga maritima MSB8
alpha-D-glucose + maltose
wild-type (alpha-D-glucose: ca. 22 mg/ml, maltose: ca. 5.5 mg/ml), mutants: W177V (alpha-D-glucose: ca. 20 mg/ml, maltose: ca. 5 mg/ml), Y178V (alpha-D-glucose: ca. 19 mg/ml, maltose: ca. 4 mg/ml), and F179V (alpha-D-glucose: ca. 17.5 mg/ml, maltose: ca. 7 mg/ml), almost no maltotriose as product for all variants, identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
3.2.1.1
starch + H2O
pH 7.0, 85°C
690543
Thermotoga maritima MSB8
alpha-D-glucose + maltose + maltotriose
mutants: H222Q (alpha-D-glucose: ca. 21 mg/ml, maltose: ca. 5 mg/ml, maltotriose: ca. 2.5 mg/ml), H222D (alpha-D-glucose: ca. 21 mg/ml, maltose: ca. 8 mg/ml, maltotriose: ca. 2.5 mg/ml), H222E (alpha-D-glucose: ca. 13 mg/ml, maltose: ca. 8 mg/ml, maltotriose: ca. 2.0 mg/ml), and V259W (alpha-D-glucose: ca. 16 mg/ml, maltose: ca. 8 mg/ml, maltotriose: ca. 2.5 mg/ml), identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
3.2.1.1
starch + methanol
-
690543
Thermotoga maritima MSB8
methylglucoside + alpha-D-glucose
methylglucoside: wild-type ca. 7.5 mg/ml, mutant H222Q ca. 13 mg/ml, mutant H222E ca. 11 mg/ml, mutant H222D ca. 11 mg/mk, identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
Synonyms
EC Number
Synonyms
Commentary
Organism
3.2.1.1
AmyA
-
Thermotoga maritima MSB8
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.2.1.1
85
-
optimum, assay at
Thermotoga maritima MSB8
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.2.1.1
85
-
the wild-type AmyA and the mutants are stable for more thant 30 h at 85°C. No loss of activity is observed
Thermotoga maritima MSB8
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.2.1.1
7
-
assay at
Thermotoga maritima MSB8
Application (protein specific)
EC Number
Application
Commentary
Organism
3.2.1.1
industry
thermal stability of the wild-type and mutant enzymes fulfill the stability requirements for industrial processes
Thermotoga maritima MSB8
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
3.2.1.1
F179V
yield a similar product profile to that of the wild-type enzyme
Thermotoga maritima MSB8
3.2.1.1
H222D
show different product spectrum than wild-type, in addition small amounts of maltotriose are produced. When methanol and butanol are used as nucleophil instead of H2O, the mutant produces more methylglucoside and butylglucoside than wild-type, respectively
Thermotoga maritima MSB8
3.2.1.1
H222E
show different product spectrum than wild-type, in addition small amounts of maltotriose are produced. When methanol is used as nucleophil instead of H2O, more methylglucoside is produced than with wild-type enzyme. Butanol produces almost equal amount in both wild-type and H222E mutant
Thermotoga maritima MSB8
3.2.1.1
H222Q
show different product spectrum than wild-type, in addition small amounts of maltotriose are produced. When methanol and butanol are used as nucleophil instead of H2O, the mutant produces more methylglucoside and butylglucoside than wild-type, respectively
Thermotoga maritima MSB8
3.2.1.1
V259W
in addition to alpha-D-glucose and maltose the product profile is broadened with small amounts of maltotriose
Thermotoga maritima MSB8
3.2.1.1
W177V
yield a similar product profile to that of the wild-type enzyme
Thermotoga maritima MSB8
3.2.1.1
Y178V
yield a similar product profile to that of the wild-type enzyme
Thermotoga maritima MSB8
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.2.1.1
wild-type and mutants H222D, H222E, H222Q, W177V, Y178V, and F179V
Thermotoga maritima MSB8
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.2.1.1
additional information
-
1000 U/mg for the wild-type enzyme and the mutants W177V, Y178V, and F179V
Thermotoga maritima MSB8
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
3.2.1.1
starch + butanol
-
690543
Thermotoga maritima MSB8
butylglucoside + alpha-D-glucose
butylglucoside: wild-type ca. 4.5 mg/ml, mutant H222Q ca. 6.5 mg/ml, mutant H222E ca. 3.5 mg/ml, mutant H222D ca. 5.5 mg/mk, identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
3.2.1.1
starch + H2O
pH 7.0, 85°C
690543
Thermotoga maritima MSB8
alpha-D-glucose + maltose
wild-type (alpha-D-glucose: ca. 22 mg/ml, maltose: ca. 5.5 mg/ml), mutants: W177V (alpha-D-glucose: ca. 20 mg/ml, maltose: ca. 5 mg/ml), Y178V (alpha-D-glucose: ca. 19 mg/ml, maltose: ca. 4 mg/ml), and F179V (alpha-D-glucose: ca. 17.5 mg/ml, maltose: ca. 7 mg/ml), almost no maltotriose as product for all variants, identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
3.2.1.1
starch + H2O
pH 7.0, 85°C
690543
Thermotoga maritima MSB8
alpha-D-glucose + maltose + maltotriose
mutants: H222Q (alpha-D-glucose: ca. 21 mg/ml, maltose: ca. 5 mg/ml, maltotriose: ca. 2.5 mg/ml), H222D (alpha-D-glucose: ca. 21 mg/ml, maltose: ca. 8 mg/ml, maltotriose: ca. 2.5 mg/ml), H222E (alpha-D-glucose: ca. 13 mg/ml, maltose: ca. 8 mg/ml, maltotriose: ca. 2.0 mg/ml), and V259W (alpha-D-glucose: ca. 16 mg/ml, maltose: ca. 8 mg/ml, maltotriose: ca. 2.5 mg/ml), identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
3.2.1.1
starch + methanol
-
690543
Thermotoga maritima MSB8
methylglucoside + alpha-D-glucose
methylglucoside: wild-type ca. 7.5 mg/ml, mutant H222Q ca. 13 mg/ml, mutant H222E ca. 11 mg/ml, mutant H222D ca. 11 mg/mk, identified and analyzed by thin-layer-chromatography and HPLC
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.2.1.1
85
-
optimum, assay at
Thermotoga maritima MSB8
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.2.1.1
85
-
the wild-type AmyA and the mutants are stable for more thant 30 h at 85°C. No loss of activity is observed
Thermotoga maritima MSB8
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.2.1.1
7
-
assay at
Thermotoga maritima MSB8