Cloned (Comment) | Organism |
---|---|
Escherichia coli DH5alphaF' grown in Luria-Bertani broth, 37°C with plasmid pTZ57R/T, expression in yeast Pichia pastoris with recombinant plasmid pPICZ-BK193 in Yeast Extract Peptone Dextrose | uncultured microorganism |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.22 | - |
pullulan | optimal pH 7.0, optimal temperature 75°C | uncultured microorganism |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | essential for enzyme activity, also tested were Mg2+, Sr2+, Zn2+, Cu2+, Fe2+, Ni2+, Mn2+ | uncultured microorganism |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
66000 | - |
SDS-PAGE, 605 amino acid product of 1815 bp gene sequence with N-terminal signal sequence of 27 amino acids (M1-Q27) | uncultured microorganism |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
uncultured microorganism | A7LI67 | gene Env Npu193A of environmental soil DNA sample from a hot spring in Thailand and expressed in yeast Pichia pastoris KM71 with plasmid pPICZalphaA | - |
Purification (Comment) | Organism |
---|---|
gene purification by agarose gel electrophoresis, enzyme purification by size exclusion chromatography, elution via Superdex 200 10/300 GL column with 50 mM MOPS, pH7, 50 mM NaCl | uncultured microorganism |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
very poor activity with substrate gamma-cyclodextrin, 50 mM MOPS buffer, pH 7.0, 2 mM CaCl2, 75°C, 45 min | uncultured microorganism |
0.15 | - |
Vmax, substrate starch, 50 mM MOPS buffer, pH 7.0, 2 mM CaCl2, 75°C, 45 min | uncultured microorganism |
23.24 | - |
Vmax, substrate pullulan, 50 mM MOPS buffer, pH 7.0, 2 mM CaCl2, 75°C, 45 min | uncultured microorganism |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
gamma-cyclodextrin + H2O | not alpha or beta-cyclodextrins | uncultured microorganism | maltose + ? | no glucose | ? | |
linear maltooligosaccharides + H2O | - |
uncultured microorganism | maltose + ? | no glucose | ? | |
pullulan + H2O | - |
uncultured microorganism | maltotriose + ? | no panose | ? | |
starch + H2O | not alpha or beta-cyclodextrins | uncultured microorganism | maltose + ? | no glucose | ? |
Synonyms | Comment | Organism |
---|---|---|
cyclomaltodextrinase | - |
uncultured microorganism |
Env Npu193A | - |
uncultured microorganism |
neopullulanase-like enzyme | - |
uncultured microorganism |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | - |
substrate pullulan: 50 mM MOPS, pH 7, 2 mM CaCl2 | uncultured microorganism |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 90 | substrate pullulan: more than 80% activity from 65-80°C, 70-80% activity from 55-60°C, about 40% activity between 40 and 50°C, about 20-30% activity at 30°C and beyond 80°C | uncultured microorganism |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | 75 | substrate pullulan, pre-incubation at 45, 50, 60, 65, 70, 75°C for 30, 60, and 120 min. about 80% activity after 120 min between 45 and 60°C, more than 60% activity after 120 min at 65°C, about 50% after 120 min at 70°C, around 20% activity after 120 min at 75°C | uncultured microorganism |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
substrate pullulan | uncultured microorganism |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 11 | substrate pullulan: at least 50% activity retained between pH 5 and 9, about 20% activity at pH 4 and beyond pH 10. buffers of 100 mM pH 4: sodium citrate, pH 5-6: acetic acid, pH 6.5-7: MOPS, pH 7-9: Tris-HCl, pH 9-11: glycine | uncultured microorganism |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
3 | 9 | substrate pullulan, 25°C, 4 h pre-incubation at pH 3: 50 mM sodium citrate, pH 5: 50 mM sodium acetate, pH 7: 50 mM Tris-HCl, pH 9: 50 mM glycin, more than 50% activity retained at pH 3, more than 80% at pH 5, about 100% at pH 7 and 9 | uncultured microorganism |