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Literature summary extracted from

  • Brown, S.H.; Kelly, R.M.
    Characterization of amylolytic enzymes, having both alpha-1,4 and alpha-1,6 hydrolytic activity, from the thermophilic archaea Pyrococcus furiosus and Thermococcus litoralis (1993), Appl. Environ. Microbiol., 59, 2614-2621.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.41 Ca2+ 5 mM, the pullulan hydrolyzing activity is stimulated by about 25%, slight enhancing effect on hydrolysis of 4-nitrophenyl-alpha-D-maltotrioside Pyrococcus furiosus
3.2.1.41 Ca2+ 5 mM, the pullulan hydrolyzing activity is stimulated by about 25%, slight enhancing effect on hydrolysis of 4-nitrophenyl-alpha-D-maltotrioside Thermococcus litoralis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.41 Ca2+ 5 mM, starch-hydrolyzing activity is inhibited by about 30%, hydrolytic activity against glycogen is slightly inhibited Pyrococcus furiosus
3.2.1.41 Ca2+ 5 mM, starch-hydrolyzing activity is inhibited by about 14%, hydrolytic activity against glycogen is slightly inhibited Thermococcus litoralis
3.2.1.41 Mg2+ reduces the thermoactivity of the enzymes below the level seen in the absence of added metal Pyrococcus furiosus
3.2.1.41 Mg2+ reduces the thermoactivity of the enzymes below the level seen in the absence of added metal Thermococcus litoralis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.41 additional information
-
 additional information Ca2+ causes a 10fold decrease in Km Pyrococcus furiosus
3.2.1.41 additional information
-
 additional information Ca2+ causes a 10fold decrease in Km Thermococcus litoralis
3.2.1.41 0.037
-
 4-nitrophenyl alpha-D-maltotrioside pH 5.6, 98°C, 5 mM Ca2+ Thermococcus litoralis
3.2.1.41 0.067
-
 4-nitrophenyl alpha-D-maltotrioside pH 5.6, 98°C, no metal added Thermococcus litoralis
3.2.1.41 0.077
-
 4-nitrophenyl alpha-D-maltotrioside pH 5.6, 98°C, 5 mM Ca2+ Pyrococcus furiosus
3.2.1.41 0.664
-
 4-nitrophenyl alpha-D-maltotrioside pH 5.6, 98°C, no metal added Pyrococcus furiosus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.41 extracellular
-
 Thermococcus litoralis
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.41 Ca2+ thermoactivity and thermostability of the enzyme is enhanced in the presence of 5 mM Ca2+, and under these conditions, enzyme activity can be measured at temperatures of up to 130 to 140°C Pyrococcus furiosus
3.2.1.41 Ca2+ thermoactivity and thermostability of the enzyme is enhanced in the presence of 5 mM Ca2+, and under these conditions, enzyme activity can be measured at temperatures of up to 130 to 140°C Thermococcus litoralis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.41 110000
-
 1 * 110000, SDS-PAGE Pyrococcus furiosus
3.2.1.41 119000
-
 1 * 119000, SDS-PAGE Thermococcus litoralis
3.2.1.41 125000
-
 gel filtration Thermococcus litoralis
3.2.1.41 135000
-
 gel filtration Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.41 Pyrococcus furiosus
-
-
-
3.2.1.41 Thermococcus litoralis
-
-
-
3.2.1.41 Thermococcus litoralis DSM 5473
-
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.2.1.41 glycoprotein
-
 Pyrococcus furiosus
3.2.1.41 glycoprotein
-
 Thermococcus litoralis

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.41
-
 Pyrococcus furiosus
3.2.1.41
-
 Thermococcus litoralis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.41 4-nitrophenyl alpha-D-maltoheptaoside + H2O hydrolyzed in an exo-fashion Pyrococcus furiosus 4-nitrophenol + alpha-D-maltoheptaose
-
 ?
3.2.1.41 4-nitrophenyl alpha-D-maltoheptaoside + H2O hydrolyzed in an exo-fashion Thermococcus litoralis 4-nitrophenol + alpha-D-maltoheptaose
-
 ?
3.2.1.41 4-nitrophenyl alpha-D-maltoheptaoside + H2O hydrolyzed in an exo-fashion Thermococcus litoralis DSM 5473 4-nitrophenol + alpha-D-maltoheptaose
-
 ?
3.2.1.41 4-nitrophenyl alpha-D-maltotrioside + H2O hydrolyzed in an exo-fashion Pyrococcus furiosus 4-nitrophenol + alpha-D-maltotriose
-
 ?
3.2.1.41 4-nitrophenyl alpha-D-maltotrioside + H2O hydrolyzed in an exo-fashion Thermococcus litoralis 4-nitrophenol + alpha-D-maltotriose
-
 ?
3.2.1.41 glycogen + H2O hydrolyzed slowly. Hydrolyzed in an endo fashion to form a series of oligosaccharides as small as glucose, with the majority of product in the DP4 to DP6 range Pyrococcus furiosus ?
-
 ?
3.2.1.41 glycogen + H2O hydrolyzed slowly. Hydrolyzed in an endo fashion to form a series of oligosaccharides as small as glucose, with the majority of product in the DP4 to DP6 range Thermococcus litoralis ?
-
 ?
3.2.1.41 glycogen + H2O hydrolyzed slowly. Hydrolyzed in an endo fashion to form a series of oligosaccharides as small as glucose, with the majority of product in the DP4 to DP6 range Thermococcus litoralis DSM 5473 ?
-
 ?
3.2.1.41 additional information hydrolyzes the alpha-1,6 linkage in pullulan, alpha-1,4 linkages in amylose and soluble starch. No activity against maltohexaose or other smaller alpha-1,4-linked oligosaccharides Pyrococcus furiosus ?
-
 ?
3.2.1.41 additional information hydrolyzes the alpha-1,6 linkage in pullulan, alpha-1,4 linkages in amylose and soluble starch. No activity against maltohexaose or other smaller alpha-1,4-linked oligosaccharides Thermococcus litoralis ?
-
 ?
3.2.1.41 additional information hydrolyzes the alpha-1,6 linkage in pullulan, alpha-1,4 linkages in amylose and soluble starch. No activity against maltohexaose or other smaller alpha-1,4-linked oligosaccharides Thermococcus litoralis DSM 5473 ?
-
 ?
3.2.1.41 pullulan + H2O higher specific activity against pullulan than against starch. Enzyme attacks only the alpha-1,6 linkages in pullulan Pyrococcus furiosus maltotriose + ?
-
 ?
3.2.1.41 pullulan + H2O higher specific activity against pullulan than against starch. Enzyme attacks only the alpha-1,6 linkages in pullulan Thermococcus litoralis maltotriose + ?
-
 ?
3.2.1.41 pullulan + H2O higher specific activity against pullulan than against starch. Enzyme attacks only the alpha-1,6 linkages in pullulan Thermococcus litoralis DSM 5473 maltotriose + ?
-
 ?
3.2.1.41 starch + H2O higher specific activity against pullulan than against starch. Hydrolyzed in an endo fashion to form a series of oligosaccharides as small as glucose, with the majority of product in the DP4 to DP6 range Pyrococcus furiosus ?
-
 ?
3.2.1.41 starch + H2O higher specific activity against pullulan than against starch. Hydrolyzed in an endo fashion to form a series of oligosaccharides as small as glucose, with the majority of product in the DP4 to DP6 range Thermococcus litoralis ?
-
 ?
3.2.1.41 starch + H2O higher specific activity against pullulan than against starch. Hydrolyzed in an endo fashion to form a series of oligosaccharides as small as glucose, with the majority of product in the DP4 to DP6 range Thermococcus litoralis DSM 5473 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.41 monomer 1 * 110000, SDS-PAGE Pyrococcus furiosus
3.2.1.41 monomer 1 * 119000, SDS-PAGE Thermococcus litoralis

Synonyms

EC Number Synonyms Comment Organism
3.2.1.41 amylopullulanase
-
 Pyrococcus furiosus
3.2.1.41 amylopullulanase
-
 Thermococcus litoralis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.41 95
-
 substrate: pullulan or starch, no metal added Thermococcus litoralis
3.2.1.41 95
-
 substrate: pullulan, no metal added Pyrococcus furiosus
3.2.1.41 115
-
 substrate: pullulan or starch, 5 mM Ca2+ Thermococcus litoralis
3.2.1.41 115
-
 substrate: starch, no metal added Pyrococcus furiosus
3.2.1.41 125
-
 substrate: pullulan or starch, 5 mM Ca2+ Pyrococcus furiosus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.41 additional information
-
 the presence of Ca2+ has a significant positive effect on enzyme activity at temperatures above 120°C, extending the range at which activity can be measured up to 130 to 140°C Pyrococcus furiosus
3.2.1.41 additional information
-
 the presence of Ca2+ has a significant positive effect on enzyme activity at temperatures above 120°C, extending the range at which activity can be measured up to 130 to 140°C Thermococcus litoralis
3.2.1.41 90 125 90°C: about 50% of maximal activity, 125°C: about 50% of maximal activity, substrate: starch, no metal added Pyrococcus furiosus
3.2.1.41 90 125 90°C: about 50% of maximal activity, 135°C: about 50% of maximal activity, substrate: pullulan, 5 mM Ca2+ Thermococcus litoralis
3.2.1.41 90 125 90°C: about 75% of maximal activity, 125°C: about 45% of maximal activity, substrate: pullulan, no metal added Pyrococcus furiosus
3.2.1.41 90 135 90°C: about 60% of maximal activity, 135°C: about 55% of maximal activity, substrate: pullulan, 5 mM Ca2+ Pyrococcus furiosus
3.2.1.41 90 115 90°C: about 60% of maximal activity, 125°C: about 70% of maximal activity, substrate: pullulan, no metal added Thermococcus litoralis
3.2.1.41 90 115 90°C: about 60% of maximal activity, 125°C: about 75% of maximal activity, substrate: starch, no metal added Thermococcus litoralis
3.2.1.41 95 125 100°C: about 65% of maximal activity, 135°C: about 80% of maximal activity, substrate: starch, 5 mM Ca2+ Thermococcus litoralis
3.2.1.41 100 135 100°C: about 40% of maximal activity, 135°C: about 60% of maximal activity, substrate: starch, 5 mM Ca2+ Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.41 additional information
-
 thermostability of the enzyme is enhanced in the presence of 5 mM Ca2+ Pyrococcus furiosus
3.2.1.41 additional information
-
 thermostability of the enzyme is enhanced in the presence of 5 mM Ca2+ Thermococcus litoralis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.41 5.5
-
-
 Pyrococcus furiosus
3.2.1.41 5.5
-
-
 Thermococcus litoralis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.41 4.5 7.8 20% of maximal activity at pH 4.5 - pH 7.8 Pyrococcus furiosus
3.2.1.41 4.5 7.8 20% of maximal activity at pH 4.5 - pH 7.8 Thermococcus litoralis