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Literature summary for 3.2.1.1 extracted from
- Characterization of an exo-acting intracellular alpha-amylase from the hyperthermophilic bacterium Thermotoga neapolitana (2010), Appl. Microbiol. Biotechnol., 86, 555-566.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | Thermotoga neapolitana |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | complete inhibition at 1 mM | Thermotoga neapolitana |  |
| Cu2+ | 52.2% residual activity at 1 mM | Thermotoga neapolitana | |
| Hg2+ | complete inhibition at 1 mM | Thermotoga neapolitana | |
| Mg2+ | 69.4% residual activity at 1 mM | Thermotoga neapolitana | |
| Mn2+ | complete inhibition at 1 mM | Thermotoga neapolitana | |
| additional information | EDTA, dithiothreitol, N-bromosuccinimide, 2-mercaptoethanol (1 mM), and SDS (2% (w/v)) have no effect on activity | Thermotoga neapolitana | |
| Urea | 25.8% residual activity at 1 mM | Thermotoga neapolitana | |
| Zn2+ | 70.4% residual activity at 1 mM | Thermotoga neapolitana | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.07 | - |
soluble starch | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 1.26 | - |
maltotriose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 1.69 | - |
maltopentaose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 2.46 | - |
4-nitrophenyl alpha-D-maltohexaoside | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 3.37 | - |
maltoheptaose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| intracellular | - |
Thermotoga neapolitana | 5622 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | Ni2+ and Ca2+ do not stimulate the activity of AmyB | Thermotoga neapolitana |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 48000 | - |
SDS-PAGE and gel filtration | Thermotoga neapolitana |
| 50120 | - |
calculated from amino acid sequence | Thermotoga neapolitana |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl alpha-D-maltohexaoside + H2O | Thermotoga neapolitana | - |
? | - |
? | |
| amylase + H2O | Thermotoga neapolitana | - |
? | - |
? | |
| amylopectin + H2O | Thermotoga neapolitana | - |
? | - |
? | |
| glycogen + H2O | Thermotoga neapolitana | - |
? | - |
? | |
| glycogen + H2O | Thermotoga neapolitana 41025 | - |
? | - |
? | |
| maltoheptaose + H2O | Thermotoga neapolitana | - |
maltose + D-glucose + ? | - |
? | |
| maltohexaose + H2O | Thermotoga neapolitana | - |
3 maltose | - |
? | |
| maltopentaose + H2O | Thermotoga neapolitana | - |
maltose + D-glucose + ? | - |
? | |
| maltotetraose + H2O | Thermotoga neapolitana | - |
2 maltose | - |
? | |
| maltotetraose + H2O | Thermotoga neapolitana 41025 | - |
2 maltose | - |
? | |
| maltotriose + H2O | Thermotoga neapolitana | - |
maltose + D-glucose | - |
? | |
| additional information | Thermotoga neapolitana | AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides | ? | - |
? | |
| additional information | Thermotoga neapolitana | inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch | ? | - |
? | |
| additional information | Thermotoga neapolitana 41025 | AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides | ? | - |
? | |
| additional information | Thermotoga neapolitana 41025 | inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch | ? | - |
? | |
| soluble starch + H2O | Thermotoga neapolitana | - |
? | - |
? | |
| soluble starch + H2O | Thermotoga neapolitana 41025 | - |
? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermotoga neapolitana | B5ARZ9 | - |
- |
| Thermotoga neapolitana 41025 | B5ARZ9 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| HiTrap SP column chromatography | Thermotoga neapolitana |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.0063 | - |
using amylopectin as substrate, in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana |
| 0.0109 | - |
using amylose as substrate, in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana |
| 0.0188 | - |
using soluble starch as substrate, in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl alpha-D-maltohexaoside + H2O | - |
Thermotoga neapolitana | ? | - |
? | |
| amylase + H2O | - |
Thermotoga neapolitana | ? | - |
? | |
| amylopectin + H2O | - |
Thermotoga neapolitana | ? | - |
? | |
| glycogen + H2O | - |
Thermotoga neapolitana | ? | - |
? | |
| glycogen + H2O | - |
Thermotoga neapolitana 41025 | ? | - |
? | |
| maltoheptaose + H2O | - |
Thermotoga neapolitana | maltose + D-glucose + ? | - |
? | |
| maltohexaose + H2O | - |
Thermotoga neapolitana | 3 maltose | - |
? | |
| maltopentaose + H2O | - |
Thermotoga neapolitana | maltose + D-glucose + ? | - |
? | |
| maltotetraose + H2O | - |
Thermotoga neapolitana | 2 maltose | - |
? | |
| maltotetraose + H2O | - |
Thermotoga neapolitana 41025 | 2 maltose | - |
? | |
| maltotriose + H2O | - |
Thermotoga neapolitana | maltose + D-glucose | - |
? | |
| additional information | AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides | Thermotoga neapolitana | ? | - |
? | |
| additional information | inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch | Thermotoga neapolitana | ? | - |
? | |
| additional information | AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides | Thermotoga neapolitana 41025 | ? | - |
? | |
| additional information | inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch | Thermotoga neapolitana 41025 | ? | - |
? | |
| soluble starch + H2O | - |
Thermotoga neapolitana | ? | - |
? | |
| soluble starch + H2O | - |
Thermotoga neapolitana 41025 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 48000, gel filtration | Thermotoga neapolitana |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AmyB | exo-acting intracellular alpha-amylase that possesses distinct characteristics from typical alpha-amylases and cyclodextrin-hydrolyzing enzymes such as maltogenic amylase, cyclodextrinase, and neopullulanase | Thermotoga neapolitana |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 75 | - |
- |
Thermotoga neapolitana |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | 100 | the enzyme retains its full activity after 7 h of incubation at 90°C, the half-life of AmyB is determined to be 50.2 and 28 h at 95°C and 100°C, respectively, thermostability is slightly enhanced by Ca2+ | Thermotoga neapolitana |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.02 | - |
soluble starch | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 25 | - |
maltoheptaose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 34 | - |
4-nitrophenyl alpha-D-maltohexaoside | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 283 | - |
maltopentaose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 809 | - |
maltotriose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
- |
Thermotoga neapolitana |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 8.5 | - |
Thermotoga neapolitana |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.286 | - |
soluble starch | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 7.5 | - |
maltoheptaose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 13.9 | - |
4-nitrophenyl alpha-D-maltohexaoside | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 167.5 | - |
maltopentaose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
| 642.1 | - |
maltotriose | in 50 mM sodium phosphate buffer (pH 6.5) at 75°C | Thermotoga neapolitana | |
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