Literature summary extracted from
Dou, S.; Chi, N.; Zhou, X.; Zhang, Q.; Pang, F.; Xiu, Z.
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)
EC Number |
Cloned (Comment) |
Organism |
---|
3.2.1.1 |
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
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
3.2.1.1 |
Co2+ |
27% inhibition at 1 mM, 68% at 10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
Cu2+ |
18% inhibition at 1 mM, 48% at 10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
EDTA |
41% inhibition at 1 mM, 79% at 10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
Fe3+ |
30% inhibition at 1 mM, 62% at 10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
Mn2+ |
14% inhibition at 1 mM, 39% at 10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
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 |
|
3.2.1.1 |
Triton X-100 |
17% inhibition at 1 mM, 51.5% at 10 mM |
Bacillus sp. dsh19-1 |
|
KM Value [mM]
EC Number |
KM Value [mM] |
KM Value Maximum [mM] |
Substrate |
Comment |
Organism |
Structure |
---|
3.2.1.1 |
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
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
3.2.1.1 |
Ca2+ |
activates 9% at 1 mM, 59% at 10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
additional information |
no or poor effects by Mg2+, K+, and Zn2+ at 1-10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
Na+ |
activates 15% at 1 mM, 61% at 10 mM |
Bacillus sp. dsh19-1 |
|
3.2.1.1 |
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
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.2.1.1 |
Bacillus sp. dsh19-1 |
A0A192GPZ8 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
3.2.1.1 |
recombinant His-tagged enzyme 8.2fold from Escherichia coli strain BL21T1R(DE3) by affinity chromatography |
Bacillus sp. dsh19-1 |
Specific Activity [micromol/min/mg]
EC Number |
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
---|
3.2.1.1 |
16.4 |
- |
purified recombinant His-tagged enzyme, pH 6.0, 20°C, substrate soluble starch |
Bacillus sp. dsh19-1 |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.2.1.1 |
soluble starch + H2O |
- |
Bacillus sp. dsh19-1 |
D-glucose + maltose + maltotriose + maltotetraose |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
3.2.1.1 |
? |
x * 50100, about, SDS-PAGE and sequence calculation |
Bacillus sp. dsh19-1 |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
3.2.1.1 |
AmyD-1 |
- |
Bacillus sp. dsh19-1 |
3.2.1.1 |
cold-active alpha-amylase |
- |
Bacillus sp. dsh19-1 |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
3.2.1.1 |
20 |
- |
recombinant enzyme |
Bacillus sp. dsh19-1 |
Temperature Range [°C]
EC Number |
Temperature Minimum [°C] |
Temperature Maximum [°C] |
Comment |
Organism |
---|
3.2.1.1 |
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]
EC Number |
Temperature Stability Minimum [°C] |
Temperature Stability Maximum [°C] |
Comment |
Organism |
---|
3.2.1.1 |
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]
EC Number |
Turnover Number Minimum [1/s] |
Turnover Number Maximum [1/s] |
Substrate |
Comment |
Organism |
Structure |
---|
3.2.1.1 |
933.5 |
- |
soluble starch |
recombinant enzyme, 20°C, pH 6.0 |
Bacillus sp. dsh19-1 |
|
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
3.2.1.1 |
6 |
- |
recombinant enzyme |
Bacillus sp. dsh19-1 |
pH Range
EC Number |
pH Minimum |
pH Maximum |
Comment |
Organism |
---|
3.2.1.1 |
4 |
10 |
over 50% of maximal activity within this range, profile overview |
Bacillus sp. dsh19-1 |
pH Stability
EC Number |
pH Stability |
pH Stability Maximum |
Comment |
Organism |
---|
3.2.1.1 |
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 |