Literature summary extracted from
Damian-Almazo, J.Y.; Moreno, A.; Lopez-Munguia, A.; Soberon, X.; Gonzalez-Munoz, F.; Saab-Rincon, G.
Enhancement of the alcoholytic activity of alpha-amylase AmyA from Thermotoga maritima MSB8 (DSM 3109) by site-directed mutagenesis (2008), Appl. Environ. Microbiol., 74, 5168-5177.
Application
EC Number |
Application |
Comment |
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 |
Protein Variants
EC Number |
Protein Variants |
Comment |
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 |
Comment |
Textmining |
---|
3.2.1.1 |
Thermotoga maritima MSB8 |
P96107 |
DSM 3109, hyperthermophilic bacterium |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
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] |
Comment |
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 |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.2.1.1 |
starch + butanol |
- |
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 |
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 |
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 |
- |
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 |
Comment |
Organism |
---|
3.2.1.1 |
AmyA |
- |
Thermotoga maritima MSB8 |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
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] |
Comment |
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 |
Comment |
Organism |
---|
3.2.1.1 |
7 |
- |
assay at |
Thermotoga maritima MSB8 |