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Literature summary for 3.2.1.1 extracted from
- A novel alpha-amylase from the cyanobacterium Nostoc sp. PCC 7119 (2010), Appl. Microbiol. Biotechnol., 86, 131-141.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| dithiothreitol | stimulates the amylolytic activity of the Amy1 protein at concentrations as low as 0.2 mM when coincubated with thioredoxin A, but is unable to activate the Amy1 protein at concentrations lower than 5 mM, the Amy1 protein activity greatly increases in the presence of dithiothreitol with a maximum (3fold)-enhanced activity at 30 mM | Anabaena sp. |  |
| thioredoxin A | the Amy1 protein activity increases linearly with increasing thioredoxin A concentrations | Anabaena sp. |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli MC1061 cells | Anabaena sp. |
General Stability
| General Stability | Organism |
|---|---|
| the activity is almost negligible with acetate and phosphate buffers at any pH value, and higher in glycine-KOH buffer than in bis-Tris propane at pH from 9.0 to 10.5, the Amy1 protein shows a significant activity in MOPS and Tris-HCl buffers. At high concentration, the protein keeps its enzymatic activity for several days | Anabaena sp. |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| alpha-cyclodextrin | the addition of alpha-cyclodextrin (0.1% w/v) induces a very weak inhibition of the Amy1 protein activity against soluble starch | Anabaena sp. | |
| ammonium sulfate | non-competitive inhibitor at concentrations higher than 20 mM | Anabaena sp. | |
| beta-cyclodextrin | the addition of beta-cyclodextrin (0.1% w/v) induces a very weak inhibition of the Amy1 protein activity against soluble starch | Anabaena sp. | |
| dithiothreitol | inhibits the enzyme activity at concentrations higher than 80 mM | Anabaena sp. | |
| gamma-cyclodextrin | the addition of gamma-cyclodextrin (0.1% w/v) induces a very weak inhibition of the Amy1 protein activity against soluble starch | Anabaena sp. | |
| maltose | non-competitive inhibitor at concentrations higher than 20 mM | Anabaena sp. | |
| additional information | beta-mercaptoethanol, mannitol, glycerol, polyethylene glycol, and Triton X-100 do not have any significant effect even at high concentration | Anabaena sp. | |
| SDS | non-reversible inactivation | Anabaena sp. | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Km is 11.0 mg/ml, using soluble potato starch as substrate, at pH 7.0 and 30°C | Anabaena sp. |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| cytoplasm | - |
Anabaena sp. | 5737 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | calcium-dependent alpha-amylase, maximum activity at 2.5 mM CaCl2 | Anabaena sp. | |
| additional information | Li+, Na+, K+, Co2+, Cu2+, Mg2+, Zn2+, and Fe3+ have no stimulatory effect on activity | Anabaena sp. |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 56000 | - |
SDS-PAGE | Anabaena sp. |
| 56700 | - |
1 * 56700, MALDI-TOF mass spectrometry | Anabaena sp. |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl maltoheptaoside + H2O | Anabaena sp. | - |
4-nitrophenol + maltoheptaoside | - |
? | |
| dextrin + H2O | Anabaena sp. | - |
? | - |
? | |
| additional information | Anabaena sp. | high-molecular-mass compounds containing alpha-1,4 linkages are the best substrates for the enzyme. After 5 min reaction, no hydrolytic activity can be detected using maltose, xylan, pullulan or alpha-, beta- and gamma-cyclodextrins as substrates | ? | - |
? | |
| oyster glycogen + H2O | Anabaena sp. | - |
? | - |
? | |
| rabbit glycogen + H2O | Anabaena sp. | - |
? | - |
? | |
| soluble potato starch + H2O | Anabaena sp. | - |
maltotriose + maltose + maltotetraose | major products of the enzymatic reaction with starch as substrate | ? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Anabaena sp. | B1VK33 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Sephadex G10 matrix gel filtration | Anabaena sp. |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.000575 | - |
crude extract, at 30°C in 10 mM MOPS buffer (pH 7.0) with 5 mM CaCl2, using 4-nitrophenyl maltoheptaoside as substrate | Anabaena sp. |
| 0.54 | - |
after 939fold purification, at 30°C in 10 mM MOPS buffer (pH 7.0) with 5 mM CaCl2, using 4-nitrophenyl maltoheptaoside as substrate | Anabaena sp. |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl maltoheptaoside + H2O | - |
Anabaena sp. | 4-nitrophenol + maltoheptaoside | - |
? | |
| dextrin + H2O | - |
Anabaena sp. | ? | - |
? | |
| additional information | high-molecular-mass compounds containing alpha-1,4 linkages are the best substrates for the enzyme. After 5 min reaction, no hydrolytic activity can be detected using maltose, xylan, pullulan or alpha-, beta- and gamma-cyclodextrins as substrates | Anabaena sp. | ? | - |
? | |
| oyster glycogen + H2O | - |
Anabaena sp. | ? | - |
? | |
| rabbit glycogen + H2O | - |
Anabaena sp. | ? | - |
? | |
| soluble potato starch + H2O | - |
Anabaena sp. | maltotriose + maltose + maltotetraose | major products of the enzymatic reaction with starch as substrate | ? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 56700, MALDI-TOF mass spectrometry | Anabaena sp. |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AMY1 | - |
Anabaena sp. |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 31 | - |
- |
Anabaena sp. |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 4 | 47 | the enzyme is relatively stable at 4°C for at least 4 h but is gradually inactivated at high temperatures, at 47°C the activity level remaining after 200 min is 25% | Anabaena sp. |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 750 | - |
oyster glycogen | at 30°C in 10 mM MOPS buffer (pH 7.0) with 5 mM CaCl2 | Anabaena sp. | |
| 1920 | - |
rabbit glycogen | at 30°C in 10 mM MOPS buffer (pH 7.0) with 5 mM CaCl2 | Anabaena sp. | |
| 1940 | - |
Dextrin | at 30°C in 10 mM MOPS buffer (pH 7.0) with 5 mM CaCl2 | Anabaena sp. | |
| 3420 | - |
soluble potato starch | at 30°C in 10 mM MOPS buffer (pH 7.0) with 5 mM CaCl2 | Anabaena sp. |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | 7.5 | - |
Anabaena sp. |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 10.5 | - |
Anabaena sp. |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Anabaena sp. | isoelectric focusing | - |
4.8 |
| Anabaena sp. | calculated from amino acid sequence | - |
5.3 |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kcat/Km is 311 s-1*mg-1*ml, using soluble potato starch as substrate, at pH 7.0 and 30°C | Anabaena sp. |
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