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Literature summary extracted from

  • Dong, G.; Vieille, C.; Zeikus, J.
    Cloning, sequencing, and expression of the gene encoding amylopullulanase from Pyrococcus furiosus and biochemical characterization of the recombinant enzyme (1997), Appl. Environ. Microbiol., 63, 3577-3584.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.41 dithiothreitol 10-50 mM, 1.2-1.3fold activation Pyrococcus furiosus
3.2.1.41 Guanidine HCl 0.1 mM, 1.1fold activation Pyrococcus furiosus
3.2.1.41 mercaptoethanol 10 mM, 1.1fold asctivation Pyrococcus furiosus
3.2.1.41 SDS 1 mM, 1.7fold activation Pyrococcus furiosus
3.2.1.41 Triton X-100 0.1-5.0%, activity on pullulan doubles after preincubation Pyrococcus furiosus
3.2.1.41 Urea 1.0-5.0 mM, 1.1fold activation Pyrococcus furiosus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.41 expressed in Escherichia coli Pyrococcus furiosus

General Stability

EC Number General Stability Organism
3.2.1.41 the enzyme is highly resistant to chemical denaturing reagents Pyrococcus furiosus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.41 Guanidine HCl 2.0 mM, about 55% inhibition Pyrococcus furiosus
3.2.1.41 SDS 5-50 mM, 25-85% inhibition Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.41 additional information
-
 pullulan Km-value for pullulan at pH 5.6, 98°C, no metal ion added: 1.6 mg/ml. Km-value for pullulan, at pH 5.6, 98°C, 0.5 mM Ca2+: 0.13 mg/ml Pyrococcus furiosus
3.2.1.41 additional information
-
 soluble starch Km-value for soluble starch at pH 5.6, 98°C, no metal ion added: 2.48 mg/ml. Km-value for soluble starch at pH 5.6, 98°C, 0.5 mM Ca2+: 1.17 mg/ml Pyrococcus furiosus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.41 Ba2+ increases activity Pyrococcus furiosus
3.2.1.41 Ca2+ increases activity and substrate affinity Pyrococcus furiosus
3.2.1.41 Co2+ increases activity Pyrococcus furiosus
3.2.1.41 Mg2+ increases activity Pyrococcus furiosus
3.2.1.41 Mn2+ increases activity Pyrococcus furiosus
3.2.1.41 Sr2+ increases activity Pyrococcus furiosus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.41 50000
-
 x * 50000, recombinant enzyme after heat treatment in denaturing buffer at 70°C or below, SDS-PAGE Pyrococcus furiosus
3.2.1.41 89000
-
-
 Pyrococcus furiosus
3.2.1.41 90000
-
 x * 90000, recombinant enzyme after heat treatment in denaturing buffer at 100°C and 110°C, SDS-PAGE Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.41 Pyrococcus furiosus O30772
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.41
-
 Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.41 amylopectin + H2O
-
 Pyrococcus furiosus ?
-
 ?
3.2.1.41 amylose + H2O
-
 Pyrococcus furiosus ?
-
 ?
3.2.1.41 glycogen + H2O
-
 Pyrococcus furiosus ?
-
 ?
3.2.1.41 additional information the enzyme also hydrolyzes oligosaccharides but much slower. The longer the oligosaccharide chain, the higher the hydrolysis rate Pyrococcus furiosus ?
-
 ?
3.2.1.41 pullulan + H2O
-
 Pyrococcus furiosus ?
-
 ?
3.2.1.41 soluble starch
-
 Pyrococcus furiosus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.41 ? x * 50000, recombinant enzyme after heat treatment in denaturing buffer at 70°C or below, SDS-PAGE Pyrococcus furiosus
3.2.1.41 ? x * 90000, recombinant enzyme after heat treatment in denaturing buffer at 100°C and 110°C, SDS-PAGE Pyrococcus furiosus

Synonyms

EC Number Synonyms Comment Organism
3.2.1.41 Apu
-
 Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.41 105
-
 with 0.5 mM Ca2+ Pyrococcus furiosus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.41 80 115 80°C: about 40% of maximal activity, 115°C: about 50% of maximal activity, with or wothout 0.5 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
-
 Ca2+ increases thermostability Pyrococcus furiosus
3.2.1.41 70
-
 the recombinant enzyme remains folded after denaturation at temperatures of below 70°C Pyrococcus furiosus
3.2.1.41 90
-
 half-life: 11 h for EDTA-treated enzyme, 44 h in presence of 5 mM Ca2+ Pyrococcus furiosus
3.2.1.41 100
-
 denaturating temperatures of above 100°C are required for complete unfolding. The enzyme remains active even after denaturation at 100 and 110°C,suggesting that unfolding isnot complete Pyrococcus furiosus

pH Optimum

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

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.41 4.3 7.2 pH 4.3: about 80% of maximal activity, pH 7.2: about 75% of maximal activity Pyrococcus furiosus