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Literature summary extracted from

  • Li, X.; Li, D.; Yin, Y.; Park, K.H.
    Characterization of a recombinant amylolytic enzyme of hyperthermophilic archaeon Thermofilum pendens with extremely thermostable maltogenic amylase activity (2010), Appl. Microbiol. Biotechnol., 85, 1821-1830.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.54 DMSO 11% activation at 10% w/v Thermofilum pendens
3.2.1.54 ethanol 13% activation at 10% w/v Thermofilum pendens
3.2.1.133 DMSO enzyme activity increases to 136% in the presence of 10% (v/v) DMSO Thermofilum pendens
3.2.1.133 ethanol enzyme activity increases to 113% in the presence of 10% (v/v) ethanol Thermofilum pendens
3.2.1.133 methanol enzyme activity increases to 104.4% in the presence of 10% (v/v) methanol Thermofilum pendens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.54 gene Tpen_1458, DNA and amino acid sequence determination and analysis, phylogenetic tree, expression as His-tagged enzyme in Escherichia coli strain MC1061 Thermofilum pendens
3.2.1.133 expressed in Escherichia coli BL21(DE3) cells Thermofilum pendens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.54 Ba2+ 23% inhibition at 5 mM Thermofilum pendens
3.2.1.54 Cu2+ 83% inhibition at 5 mM Thermofilum pendens
3.2.1.54 Fe2+ 81.6% inhibition at 5 mM Thermofilum pendens
3.2.1.54 Hg2+ 82.5% inhibition at 5 mM Thermofilum pendens
3.2.1.54 Mn2+ 50% inhibition at 5 mM Thermofilum pendens
3.2.1.54 Zn2+ 82.2% inhibition at 5 mM Thermofilum pendens
3.2.1.133 Ba2+ enzyme activity decreases to 77.3% in the presence of 5 mM BaCl2 Thermofilum pendens
3.2.1.133 Cu2+ enzyme activity decreases to 17% in the presence of 5 mM CuCl2 Thermofilum pendens
3.2.1.133 Fe2+ enzyme activity decreases to 18.4% in the presence of 5 mM FeSO4 Thermofilum pendens
3.2.1.133 Hg2+ enzyme activity decreases to 17.5% in the presence of 5 mM HgCl2 Thermofilum pendens
3.2.1.133 Mn2+ enzyme activity decreases to 50.2% in the presence of 5 mM MnCl2 Thermofilum pendens
3.2.1.133 Zn2+ enzyme activity decreases to 17.8% in the presence of 5 mM ZnCl2 Thermofilum pendens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.54 Al3+ activates 46.8% at 5 mM Thermofilum pendens
3.2.1.54 Ca2+ activates 36% at 5 mM Thermofilum pendens
3.2.1.54 Co2+ 6% activation at 5 mM Thermofilum pendens
3.2.1.54 Mg2+ activates 30% at 5 mM Thermofilum pendens
3.2.1.133 Al3+ enzyme activity increases to 146.8% in the presence of 5 mM AlCl3 Thermofilum pendens
3.2.1.133 Ca2+ enzyme activity increases to 136% in the presence of 5 mM CaCl2 Thermofilum pendens
3.2.1.133 Mg2+ enzyme activity increases to 130% in the presence of 5 mM CaCl2 Thermofilum pendens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.133 74000
-
 x * 74000, SDS-PAGE Thermofilum pendens
3.2.1.133 74252
-
 x * 74252, calculated from amino acid sequence Thermofilum pendens

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.2.1.133 DMSO enzyme activity increases to 136% in the presence of 10% (v/v) DMSO Thermofilum pendens
3.2.1.133 Ethanol enzyme activity increases to 113% in the presence of 10% (v/v) ethanol Thermofilum pendens
3.2.1.133 Methanol enzyme activity increases to 104.4% in the presence of 10% (v/v) methanol Thermofilum pendens

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.54 Thermofilum pendens
-
-
-
3.2.1.133 Thermofilum pendens
-
-
-
3.2.1.133 Thermofilum pendens Hrk 5 (DSM 2475)
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.54 recombinant His-tagged enzyme 3.0fold from Escherichia coli by heat treatment and nickel affinity chromatography Thermofilum pendens
3.2.1.133 Ni-NTA column chromatography Thermofilum pendens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.54 177
-
 95°C, pH 5.5, crude recombinant enzyme in Escherichia coli cell extract, substrate is gamma-cyclodextrin Thermofilum pendens
3.2.1.54 535.1
-
 95°C, pH 5.5, purified recombinant enzyme, substrate is gamma-cyclodextrin Thermofilum pendens
3.2.1.133 91
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM CuCl2 Thermofilum pendens
3.2.1.133 93.6
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM HgCl2 Thermofilum pendens
3.2.1.133 95.2
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM ZnCl2 Thermofilum pendens
3.2.1.133 98.5
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM FeSO4 Thermofilum pendens
3.2.1.133 177.4
-
 cell extract, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5) Thermofilum pendens
3.2.1.133 268.6
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM MnCl2 Thermofilum pendens
3.2.1.133 413.6
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM BaCl2 Thermofilum pendens
3.2.1.133 535.1
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5) Thermofilum pendens
3.2.1.133 558.6
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 10% (v/v) methanol Thermofilum pendens
3.2.1.133 568.8
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM CoCl2 Thermofilum pendens
3.2.1.133 592.9
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 10% (v/v) DMSO Thermofilum pendens
3.2.1.133 604.7
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 10% (v/v) ethanol Thermofilum pendens
3.2.1.133 695.6
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM MgCl2 Thermofilum pendens
3.2.1.133 728.8
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM CaCl2 Thermofilum pendens
3.2.1.133 785.5
-
 after 3fold purification, using gamma-cyclodextrin as substrate, at 90°C in 50 mM sodium acetate buffer (pH 5.5), with 5 mM AlCl3 Thermofilum pendens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.54 alpha-cyclodextrin + H2O 65% of the activity with gamma-cyclodextrin Thermofilum pendens maltose + D-glucose
-
 ?
3.2.1.54 alpha-cyclodextrin + H2O 65% of the activity with gamma-cyclodextrin Thermofilum pendens Hrk 5 (DSM 2475) maltose + D-glucose
-
 ?
3.2.1.54 beta-cyclodextrin + H2O 78% of the activity with gamma-cyclodextrin Thermofilum pendens maltose + D-glucose
-
 ?
3.2.1.54 beta-cyclodextrin + H2O 78% of the activity with gamma-cyclodextrin Thermofilum pendens Hrk 5 (DSM 2475) maltose + D-glucose
-
 ?
3.2.1.54 gamma-cyclodextrin + H2O during the initial stage of the reaction, only one alpha-1,4-glycosidic linkage of every gamma-cyclodexrtrin is cleaved, and then the linear maltooctaose is formed. Until almost all gamma-cyclodextrin has disappeared, maltooctaose is not further hydrolyzed. Selective degradation of gamma-cyclodextrin is preferred by the enzyme in the presence of maltooctaose. After all rings are opened at one single glycosidic linkage of cyclodextrins, the corresponding maltooligosaccharides started to be hydrolyzed into smaller maltooligosaccharides. The final hydrolysis products are maltose and glucose Thermofilum pendens maltose + D-glucose
-
 ?
3.2.1.54 maltoheptaose + H2O 24% of the activity with gamma-cyclodextrin Thermofilum pendens maltose + D-glucose + ?
-
 ?
3.2.1.54 maltoheptaose + H2O 24% of the activity with gamma-cyclodextrin Thermofilum pendens Hrk 5 (DSM 2475) maltose + D-glucose + ?
-
 ?
3.2.1.54 maltohexaose + H2O 14% of the activity with gamma-cyclodextrin Thermofilum pendens maltose + D-glucose + ?
-
 ?
3.2.1.54 maltopentaose + H2O 11% of the activity with gamma-cyclodextrin Thermofilum pendens maltose + D-glucose + ?
-
 ?
3.2.1.54 maltotetraose + H2O 6% of the activity with gamma-cyclodextrin Thermofilum pendens maltose + 2 D-glucose
-
 ?
3.2.1.54 maltotetraose + H2O 6% of the activity with gamma-cyclodextrin Thermofilum pendens Hrk 5 (DSM 2475) maltose + 2 D-glucose
-
 ?
3.2.1.54 maltotriose + H2O 5.3% of the activity with gamma-cyclodextrin Thermofilum pendens maltose + D-glucose
-
 ?
3.2.1.54 maltotriose + H2O 5.3% of the activity with gamma-cyclodextrin Thermofilum pendens Hrk 5 (DSM 2475) maltose + D-glucose
-
 ?
3.2.1.133 acarbose + H2O
-
 Thermofilum pendens acarviosine-glucose + D-glucose
-
 ?
3.2.1.133 alpha-cyclodextrin + H2O 64.7% activity compared to gamma-cyclodextrin Thermofilum pendens ?
-
 ?
3.2.1.133 beta-cyclodextrin + H2O 78.1% activity compared to gamma-cyclodextrin Thermofilum pendens ?
-
 ?
3.2.1.133 gamma-cyclodextrin + H2O maximal activity (100%) Thermofilum pendens alpha-maltose + alpha-D-glucose
-
 ?
3.2.1.133 additional information maltooligosaccharides G3-G7 show 5.4-24.1% relative activity compared to gamma-cyclodextrin Thermofilum pendens ?
-
 ?
3.2.1.133 pullulan + H2O
-
 Thermofilum pendens panose + ?
-
 ?
3.2.1.133 starch + H2O
-
 Thermofilum pendens alpha-maltose + ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.133 ? x * 74000, SDS-PAGE Thermofilum pendens
3.2.1.133 ? x * 74252, calculated from amino acid sequence Thermofilum pendens

Synonyms

EC Number Synonyms Comment Organism
3.2.1.133 maltogenic amylase
-
 Thermofilum pendens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.54 95
-
-
 Thermofilum pendens
3.2.1.133 95
-
-
 Thermofilum pendens

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.54 80 105 80°C: about 50% of maximal activity, 105°C: about 50% of maximal activity Thermofilum pendens
3.2.1.133 80 105 at 105°C, almost 50% of the maximum activity is detected Thermofilum pendens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.54 85
-
 160 min, about 50% loss of activity Thermofilum pendens
3.2.1.54 90
-
 160 min, about 45% loss of activity Thermofilum pendens
3.2.1.54 95
-
 80 min, about 55% loss of activity Thermofilum pendens
3.2.1.54 100
-
 30 min, about 60% loss of activity Thermofilum pendens
3.2.1.133 85 100 purified enzyme is extremely thermostable with a half-life of 60 min at an optimal temperature of 95°C. The enzyme retains about 80% relative activity after 60 min of incubation at 85°C and 90°C, about 50% relative activity after 60 min of incubation at 95°C, and about 25% relative activity after 60 min of incubation at 100°C Thermofilum pendens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.54 5.5
-
 substrate: gamma-cyclodextrin Thermofilum pendens
3.2.1.133 5.5
-
-
 Thermofilum pendens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.54 5 6.5 pH 5.0: about 80% of maximal activity, pH 6.5: about 50% of maximal activity Thermofilum pendens
3.2.1.133 5 6.5 more than 50% of the maximum activity is retained in the range between pH 5.0 and pH 6.5 Thermofilum pendens