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Literature summary for 3.2.1.1 extracted from
- Molecular cloning, expression, and biochemical characterization of a novel cold-active alpha-amylase from Bacillus sp. dsh19-1 (2018), Extremophiles, 22, 739-749 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21T1R(DE3) with molecular chaperones plasmid pGro7 required for enzyme solubility, subcloning in Escherichia coli strain JM109 | Bacillus sp. dsh19-1 |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | 27% inhibition at 1 mM, 68% at 10 mM | Bacillus sp. dsh19-1 |  |
| Cu2+ | 18% inhibition at 1 mM, 48% at 10 mM | Bacillus sp. dsh19-1 | |
| EDTA | 41% inhibition at 1 mM, 79% at 10 mM | Bacillus sp. dsh19-1 | |
| Fe3+ | 30% inhibition at 1 mM, 62% at 10 mM | Bacillus sp. dsh19-1 | |
| Mn2+ | 14% inhibition at 1 mM, 39% at 10 mM | Bacillus sp. dsh19-1 | |
| NaCl | enzyme AmyD-1 displays extreme salt tolerance, with the highest activity in the presence of 2.0 M NaCl and 60.5% of activity in 5.0 M NaCl, 40% inhibition at 5 M | Bacillus sp. dsh19-1 | |
| Triton X-100 | 17% inhibition at 1 mM, 51.5% at 10 mM | Bacillus sp. dsh19-1 | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics, Km and Vmax of AmyD-1 in 2.0 M NaCl are 2.8 mg/ml and 21.8 mg*ml/ml, respectively, at 20°C and pH 6.0 | Bacillus sp. dsh19-1 |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | activates 9% at 1 mM, 59% at 10 mM | Bacillus sp. dsh19-1 | |
| additional information | no or poor effects by Mg2+, K+, and Zn2+ at 1-10 mM | Bacillus sp. dsh19-1 | |
| Na+ | activates 15% at 1 mM, 61% at 10 mM | Bacillus sp. dsh19-1 | |
| NaCl | enzyme AmyD-1 displays extreme salt tolerance, with the highest activity in the presence of 2.0 M NaCl and 60.5% of activity in 5.0 M NaCl, 40% inhibition at 5 M | Bacillus sp. dsh19-1 | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus sp. dsh19-1 | A0A192GPZ8 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme 8.2fold from Escherichia coli strain BL21T1R(DE3) by affinity chromatography | Bacillus sp. dsh19-1 |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 16.4 | - |
purified recombinant His-tagged enzyme, pH 6.0, 20°C, substrate soluble starch | Bacillus sp. dsh19-1 |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| soluble starch + H2O | - |
Bacillus sp. dsh19-1 | D-glucose + maltose + maltotriose + maltotetraose | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 50100, about, SDS-PAGE and sequence calculation | Bacillus sp. dsh19-1 |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AmyD-1 | - |
Bacillus sp. dsh19-1 |
| cold-active alpha-amylase | - |
Bacillus sp. dsh19-1 |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 20 | - |
recombinant enzyme | Bacillus sp. dsh19-1 |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 4 | 70 | about 35.7% of maximal activity at 4°C, about 50% at 10°C and 45°C, about 10% at 65°C, inactivation at 75°C | Bacillus sp. dsh19-1 |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 10 | 45 | purified recombinant His-tagged enzyme, over 50% activity remaining, completely stable at 20°C, over 80% activity remaining at 15-35°C | Bacillus sp. dsh19-1 |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 933.5 | - |
soluble starch | recombinant enzyme, 20°C, pH 6.0 | Bacillus sp. dsh19-1 |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
recombinant enzyme | Bacillus sp. dsh19-1 |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 4 | 10 | over 50% of maximal activity within this range, profile overview | Bacillus sp. dsh19-1 |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 4 | 9 | purified recombinant His-tagged enzyme, over 50% activity remaining, completely stable at pH 6.0, over 80% activity remaining at pH 5.0-8.0 | Bacillus sp. dsh19-1 |
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