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Literature summary for 3.2.1.1 extracted from
- Strain improvement studies on Microbacterium foliorum GA2 for production of alpha-amylase in solid state fermentation Biochemical characteristics and wash performance analysis at low temperatures (2018), J. Gen. Appl. Microbiol., 63, 347-354 .
Application
| Application | Comment | Organism |
|---|---|---|
| additional information | cold-active alpha-amylase from mutant Microbacterium foliorum strain GA2 can be profitably used in detergents for laundry and automatic dishwashing to degrade the residues of starchy foods | Microbacterium foliorum |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | random chemical mutagenesis by the treatment of cells with sodium azide, alkali-tolerant Microbacterium foliorum strain GA2 is randomly mutated using UV radiation and sodium azide to obtain a mutant with higher cold-active extracellular amylolytic activity, designated as MFSD20, which is selected owing to its higher amylase activity at 20°C. Under optimized conditions, amylase production is achieved best with raw banana peels (5000 units) in solid-state fermentation (SSF) | Microbacterium foliorum |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | 47% inhibition of wild-type and mutant enzymes at 5 mM | Microbacterium foliorum |  |
| Cu2+ | 62% inhibition of wild-type and mutant enzymes at 5 mM | Microbacterium foliorum | |
| Fe2+ | strong inhibition of wild-type and mutant enzymes at 5 mM | Microbacterium foliorum | |
| Hg2+ | 73% inhibition of wild-type and mutant enzymes at 5 mM | Microbacterium foliorum | |
| Zn2+ | 54% inhibition of wild-type and mutant enzymes at 5 mM | Microbacterium foliorum | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | apparent Km and Vmax values for the hydrolysis of soluble starch are 4 mg/ml and 1111 units/mg for the wild-type enzyme, and 2 mg/ml and 844.5 units/mg for the mutant enzyme, respectively | Microbacterium foliorum |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | the enzyme is secreted | Microbacterium foliorum | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | highly activating at 5 mM | Microbacterium foliorum | |
| additional information | no effect by Mg2+ at 5 mM | Microbacterium foliorum |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| starch + H2O | Microbacterium foliorum | - |
? | - |
? | |
| starch + H2O | Microbacterium foliorum GA2 | - |
? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Microbacterium foliorum | A0A1S7D3W6 | - |
- |
| Microbacterium foliorum GA2 | A0A1S7D3W6 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| extracellular native wild-type and mutant enzymes 14.2fold and 19.6fold, respectively, from culture supernatants by dialysis, ammonium sulfate fractionation, dialysis, and anion exchange chromatography, to homogeneity | Microbacterium foliorum |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| additional information | production of alpha-amylase in solid state fermentation from mutated strain | Microbacterium foliorum | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
one unit of enzyme activity is defined as the quantity of enzyme that causes a 0.01% reduction of absorbance at 690 nm of starch-iodine solution at 20°C in one min per ml | Microbacterium foliorum |
| 595.5 | - |
purified native wild-type enzyme, pH 5.0, 20°C | Microbacterium foliorum |
| 700 | - |
purified native mutant enzyme, pH 5.0, 20°C | Microbacterium foliorum |
| 804.88 | - |
purified native wild-type enzyme, pH 8.0, 20°C | Microbacterium foliorum |
| 1022.36 | - |
purified native mutant enzyme, pH 5.0, 4°C | Microbacterium foliorum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| starch + H2O | - |
Microbacterium foliorum | ? | - |
? | |
| starch + H2O | - |
Microbacterium foliorum GA2 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 66000, wild-type and mutant enzymes, SDS-PAGE | Microbacterium foliorum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| cold-active alpha-amylase | - |
Microbacterium foliorum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 4 | - |
mutant MFSD20 | Microbacterium foliorum |
| 15 | - |
wild-type enzyme | Microbacterium foliorum |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 4 | 45 | purified cold-active alpha-amylase is active at 4-45°C. Optimum temperature is 15°C, 84% of maximal activity at 4°C and 25°C, 62% at 35°C, rapid drop to only 5% of maximal activity at 45°C. Temperature optimum of mutagenized alpha-amylase from the MFSZ20 strain shifts to 4°C (1022.36 units/mg), 92% of maximal activity at 25°C, 87% at 35°C, and 17% at 45°C | Microbacterium foliorum |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 20 | - |
purified wild-type enzyme shows 64% activity remaining after 2 h, the mutant enzyme from strain MFSD20 shows 70% remaining activity after 2 h | Microbacterium foliorum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
wild-type enzyme | Microbacterium foliorum |
| 10 | - |
mutant enzyme MFSD20 | Microbacterium foliorum |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 12 | purified wild-type alpha-amylase shows good activity over a broad pH range (pH 7-10) and the optimum pH for enzyme activity is pH 8.0. The specific activity is 512.27, 599.65, 773.29 and 757.5 units/mg at pH 5.0, 6.0, 7.0, and 9.0, respectively. Specific activity is still 694.43 units (86%) at pH 10. But beyond pH 10, enzymatic activity is drastically reduced and only 536.9 units/mg specific activity is observed at pH 11.0 which further drops to 157.72 units/mg at pH 12.0 | Microbacterium foliorum |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | both purified wild-type enzyme and mutant enzyme (from strain MFSD20) show very high stability with different detergents at 20°C, overview | Microbacterium foliorum |
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