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Literature summary for 3.2.1.1 extracted from
- Purification, characterization, and partial primary sequence of a major-maltotriose-producing alpha-amylase, ScAmy43, from Sclerotinia sclerotiorum (2008), J. Microbiol. Biotechnol., 18, 1555-1563.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | 5 mM, pH 4.0, 55°C, 172% relative activity, strongly increase the activity, probably due to a better accessibility of the substrate to the catalytic site after disruption of the intersulfide bridge | Sclerotinia sclerotiorum |  |
| dithiothreitol | 5 mM, pH 4.0, 55°C, 166% relative activity, strongly increase the activity, probably due to a better accessibility of the substrate to the catalytic site after disruption of the intersulfide bridge | Sclerotinia sclerotiorum | |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| BaCl2 | 5 mM, pH 4.0, 55°C, 25% relative activity | Sclerotinia sclerotiorum | |
| CuCl2 | 5 mM, pH 4.0, 55°C, 26% relative activity | Sclerotinia sclerotiorum | |
| EDTA | 5, 10, and 20 mM, pH 4.0, 55°C, 95% relative activity, enzyme is not sensitive to chelating agent, probably not a metalloenzyme | Sclerotinia sclerotiorum | |
| EGTA | 5, 10, and 20 mM, pH 4.0, 55°C, 100% relative activity, enzyme is not sensitive to chelating agent, probably not a metalloenzyme | Sclerotinia sclerotiorum | |
| FeCl2 | 5 mM, pH 4.0, 55°C, 85% relative activity | Sclerotinia sclerotiorum | |
| MnCl2 | 5 mM, pH 4.0, 55°C, 44% relative activity | Sclerotinia sclerotiorum | |
| SDS | 2%, pH 4.0, 55°C, 11% relative activity | Sclerotinia sclerotiorum | |
| Urea | 5 M, pH 4.0, 55°C, 63% relative activity | Sclerotinia sclerotiorum | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
soluble starch | Km: 1.66 mg/ml, Vmax: 0.0001 mmol/min/ml | Sclerotinia sclerotiorum |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| CaCl2 | 5 mM, pH 4.0, 55°C, 111% relative activity | Sclerotinia sclerotiorum | |
| MgCl2 | 5 mM, pH 4.0, 55°C, 103% relative activity | Sclerotinia sclerotiorum | |
| ZnCl2 | 5 mM, pH 4.0, 55°C, 104% relative activity | Sclerotinia sclerotiorum | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 40000 | - |
gel filtration | Sclerotinia sclerotiorum |
| 43000 | - |
SDS-PAGE | Sclerotinia sclerotiorum |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Sclerotinia sclerotiorum | - |
pathogenic fungus, grown on oats flour | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Sclerotinia sclerotiorum |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| culture medium | - |
Sclerotinia sclerotiorum | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | ScAmy43 does not hydrolyze native starches | Sclerotinia sclerotiorum | ? | - |
? | |
| starch + H2O | pH 4, 55°C, various starches are tested: soluble potato starch as reference 100% relative activity, gelatinized amylose: 74%, potato native starch: 0.2%, potato gelatinized starch: 106%, maize native starch: 0.8%, maize gelatinized starch: 53%, rice native starch: 0.5%, and rice gelatinized starch: 52% | Sclerotinia sclerotiorum | maltotriose + glucose + maltose + maltotetraose | identified by thin-layer-chromatography, maltotriose represents 70% of the end products, only traces of glucose, small amounts of maltose and maltotetraose | ? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 43000, SDS-PAGE | Sclerotinia sclerotiorum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| maltotriose-producing alpha-amylase | - |
Sclerotinia sclerotiorum |
| ScAmy43 | - |
Sclerotinia sclerotiorum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
assay at | Sclerotinia sclerotiorum |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | 65 | 40°C: ca. 40% relative activity, 65°C: ca. 40% relative activity, pH 4.0 for 30 min | Sclerotinia sclerotiorum |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
stable in the absence of substrate during 30 min pre-incubation, assay at pH 4.0 and 55°C, above 40°C rapid inactivation | Sclerotinia sclerotiorum |
| 50 | - |
in the presence of 1% (w/v) potato starch the thermostability is enhanced above 50°C | Sclerotinia sclerotiorum |
| 55 | - |
20% of maximal activity after 30 min pre-incubation in the absence of a substrate, but the enzyme is stable during short time assay for 30 min | Sclerotinia sclerotiorum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 4 | - |
- |
Sclerotinia sclerotiorum |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 3 | 6.5 | pH 3: below 10%, pH 6.5: ca. 10 relativ activity, 50% of maximal activity at pH 3.5 and 5.5, 55°C | Sclerotinia sclerotiorum |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 4 | 7 | ScAmy43 is stable over a narrow range, pH 4.0: below 60% residual activity, pH 7: below 80% residual activity, after incubation at various pH at 4°C for 24 h, assay at pH 4.0 and 55°C for 30 min | Sclerotinia sclerotiorum |
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