Literature summary for 3.2.1.1 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.

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Application

Application	Comment	Organism
industry	thermal stability of the wild-type and mutant enzymes fulfill the stability requirements for industrial processes	Thermotoga maritima MSB8

Protein Variants

Protein Variants	Comment	Organism
F179V	yield a similar product profile to that of the wild-type enzyme	Thermotoga maritima MSB8
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
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
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
V259W	in addition to alpha-D-glucose and maltose the product profile is broadened with small amounts of maltotriose	Thermotoga maritima MSB8
W177V	yield a similar product profile to that of the wild-type enzyme	Thermotoga maritima MSB8
Y178V	yield a similar product profile to that of the wild-type enzyme	Thermotoga maritima MSB8

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima MSB8	P96107	DSM 3109, hyperthermophilic bacterium	-

Purification (Commentary)

Purification (Comment)	Organism
wild-type and mutants H222D, H222E, H222Q, W177V, Y178V, and F179V	Thermotoga maritima MSB8

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	1000 U/mg for the wild-type enzyme and the mutants W177V, Y178V, and F179V	Thermotoga maritima MSB8

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
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	?
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	?
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	?
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

Synonyms	Comment	Organism
AmyA	-	Thermotoga maritima MSB8

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
85	-	optimum, assay at	Thermotoga maritima MSB8

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Thermotoga maritima MSB8

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Release 2023.1 (January 2023)