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Literature summary for 3.2.1.41 extracted from
- 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.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| dithiothreitol | 10-50 mM, 1.2-1.3fold activation | Pyrococcus furiosus |  |
| Guanidine HCl | 0.1 mM, 1.1fold activation | Pyrococcus furiosus | |
| mercaptoethanol | 10 mM, 1.1fold asctivation | Pyrococcus furiosus | |
| SDS | 1 mM, 1.7fold activation | Pyrococcus furiosus | |
| Triton X-100 | 0.1-5.0%, activity on pullulan doubles after preincubation | Pyrococcus furiosus | |
| Urea | 1.0-5.0 mM, 1.1fold activation | Pyrococcus furiosus | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli | Pyrococcus furiosus |
General Stability
| General Stability | Organism |
|---|---|
| the enzyme is highly resistant to chemical denaturing reagents | Pyrococcus furiosus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Guanidine HCl | 2.0 mM, about 55% inhibition | Pyrococcus furiosus | |
| SDS | 5-50 mM, 25-85% inhibition | Pyrococcus furiosus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 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 | |
| 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
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ba2+ | increases activity | Pyrococcus furiosus | |
| Ca2+ | increases activity and substrate affinity | Pyrococcus furiosus | |
| Co2+ | increases activity | Pyrococcus furiosus | |
| Mg2+ | increases activity | Pyrococcus furiosus | |
| Mn2+ | increases activity | Pyrococcus furiosus | |
| Sr2+ | increases activity | Pyrococcus furiosus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 50000 | - |
x * 50000, recombinant enzyme after heat treatment in denaturing buffer at 70°C or below, SDS-PAGE | Pyrococcus furiosus |
| 89000 | - |
- |
Pyrococcus furiosus |
| 90000 | - |
x * 90000, recombinant enzyme after heat treatment in denaturing buffer at 100°C and 110°C, SDS-PAGE | Pyrococcus furiosus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pyrococcus furiosus | O30772 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Pyrococcus furiosus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| amylopectin + H2O | - |
Pyrococcus furiosus | ? | - |
? | |
| amylose + H2O | - |
Pyrococcus furiosus | ? | - |
? | |
| glycogen + H2O | - |
Pyrococcus furiosus | ? | - |
? | |
| additional information | the enzyme also hydrolyzes oligosaccharides but much slower. The longer the oligosaccharide chain, the higher the hydrolysis rate | Pyrococcus furiosus | ? | - |
? | |
| pullulan + H2O | - |
Pyrococcus furiosus | ? | - |
? | |
| soluble starch | - |
Pyrococcus furiosus | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 50000, recombinant enzyme after heat treatment in denaturing buffer at 70°C or below, SDS-PAGE | Pyrococcus furiosus |
| ? | x * 90000, recombinant enzyme after heat treatment in denaturing buffer at 100°C and 110°C, SDS-PAGE | Pyrococcus furiosus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Apu | - |
Pyrococcus furiosus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 105 | - |
with 0.5 mM Ca2+ | Pyrococcus furiosus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 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]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
Ca2+ increases thermostability | Pyrococcus furiosus |
| 70 | - |
the recombinant enzyme remains folded after denaturation at temperatures of below 70°C | Pyrococcus furiosus |
| 90 | - |
half-life: 11 h for EDTA-treated enzyme, 44 h in presence of 5 mM Ca2+ | Pyrococcus furiosus |
| 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
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | - |
- |
Pyrococcus furiosus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 4.3 | 7.2 | pH 4.3: about 80% of maximal activity, pH 7.2: about 75% of maximal activity | Pyrococcus furiosus |
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