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Literature summary for 3.2.1.1 extracted from
- Extensive hydrolysis of raw rice starch by a chimeric alpha-amylase engineered with alpha-amylase (AmyP) and a starch-binding domain from Cryptococcus sp. S-2 (2018), Appl. Microbiol. Biotechnol., 102, 743-750 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene Amy-1E, recombinant expression of C-terminally His-tagged wild-type enzyme AmyP and chimeric mutant enzyme AmyP-Cr in Escherichia coli | uncultured bacterium |
| gene amy-CS2, recombinant expression of C-terminally His-tagged chimeric mutant enzyme AmyP-Cr in Escherichia coli | Cryptococcus sp. S-2 |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | construction of a recombinant chimeric alpha-amylase (AmyP-Cr) by a catalytic core of alpha-amylase (AmyP) from a marine metagenomic library and a starch-binding domain (SBDCr) of alpha-amylase from Cryptococcus sp. S-2. The molecular fusion does not alter optimum pH, optimum temperature, hydrolysis products, and an ability of preferential and rapid degradation towards raw rice starch, but catalytic efficiency and thermostability are remarkably improved compared with those of the wild-type AmyP. The chimeric AmyP-Cr achieves the final hydrolysis degree of 61.7% for 10% raw rice starch and 47.3% for 15% raw rice starch after 4 h at 40°C with 1.0 U per mg of raw starch, the catalytic efficiency is 3.6-4.0 times higher than that of wild-type AmyP | uncultured bacterium |
| additional information | construction of a recombinant chimeric alpha-amylase (AmyP-Cr) by a catalytic core of alpha-amylase (AmyP) from a marine metagenomic library and a starch-binding domain (SBDCr) of alpha-amylase from Cryptococcus sp. S-2. The molecular fusion does not alter optimum pH, optimum temperature, hydrolysis products, and an ability of preferential and rapid degradation towards raw rice starch, but catalytic efficiency and thermostability are remarkably improved compared with those of the wild-type AmyP. The chimeric AmyP-Cr achieves the final hydrolysis degree of 61.7% for 10% raw rice starch and 47.3% for 15% raw rice starch after 4 h at 40°C with 1.0 U per mg of raw starch, the catalytic efficiency is 3.6-4.0 times higher than that of wild-type AmyP | Cryptococcus sp. S-2 |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | required | uncultured bacterium |  |
| Ca2+ | required | Cryptococcus sp. S-2 | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| starch + H2O | uncultured bacterium | - |
? | - |
? | |
| starch + H2O | Cryptococcus sp. S-2 | - |
? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Cryptococcus sp. S-2 | Q92394 | - |
- |
| uncultured bacterium | D9MZ14 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged chimeric mutant enzyme AmyP-Cr from Escherichia coli by nickel affinity chromatography | Cryptococcus sp. S-2 |
| recombinant His-tagged chimeric wild-type and mutant mutant enzymes from Escherichia coli by nickel affinity chromatography | uncultured bacterium |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 7.4 | - |
purified recombinant His-tagged wild-type enzyme AmyP, pH 7.5, 40°C, substrate wheat starch | uncultured bacterium |
| 8.9 | - |
purified recombinant His-tagged wild-type enzyme AmyP, pH 7.5, 40°C, substrate pea starch | uncultured bacterium |
| 13.3 | - |
purified recombinant His-tagged wild-type enzyme AmyP, pH 7.5, 40°C, substrate potato starch | uncultured bacterium |
| 14.6 | - |
purified recombinant His-tagged chimeric mutant enzyme AmyP-Cr, pH 7.5, 40°C, substrate pea starch | uncultured bacterium |
| 17.9 | - |
purified recombinant His-tagged wild-type enzyme AmyP, pH 7.5, 40°C, substrate mung bean starch | uncultured bacterium |
| 25.6 | - |
purified recombinant His-tagged chimeric mutant enzyme AmyP-Cr, pH 7.5, 40°C, substrate potato starch | uncultured bacterium |
| 29.4 | - |
purified recombinant His-tagged chimeric mutant enzyme AmyP-Cr, pH 7.5, 40°C, substrate mung bean starch | uncultured bacterium |
| 33.2 | - |
purified recombinant His-tagged chimeric mutant enzyme AmyP-Cr, pH 7.5, 40°C, substrate wheat starch | uncultured bacterium |
| 39.6 | - |
purified recombinant His-tagged wild-type enzyme AmyP, pH 7.5, 40°C, substrate maize starch | uncultured bacterium |
| 148.6 | - |
purified recombinant His-tagged wild-type enzyme AmyP, pH 7.5, 40°C, substrate raw rice starch | uncultured bacterium |
| 175.8 | - |
purified recombinant His-tagged chimeric mutant enzyme AmyP-Cr, pH 7.5, 40°C, substrate maize starch | uncultured bacterium |
| 426.7 | - |
purified recombinant His-tagged chimeric mutant enzyme AmyP-Cr, pH 7.5, 40°C | Cryptococcus sp. S-2 |
| 426.7 | - |
purified recombinant His-tagged chimeric mutant enzyme AmyP-Cr, pH 7.5, 40°C, substrate raw rice starch | uncultured bacterium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| maize starch + H2O | - |
uncultured bacterium | ? | - |
? | |
| mung bean starch + H2O | - |
uncultured bacterium | ? | - |
? | |
| pea starch + H2O | - |
uncultured bacterium | ? | - |
? | |
| potato starch + H2O | - |
uncultured bacterium | ? | - |
? | |
| raw rice starch + H2O | recombinant chimeric enzyme mutant | Cryptococcus sp. S-2 | ? | - |
? | |
| raw rice starch + H2O | wild-type enzyme AmyP recombinant chimeric enzyme mutant | uncultured bacterium | D-glucose + maltose + maltotriose | - |
? | |
| starch + H2O | - |
uncultured bacterium | ? | - |
? | |
| starch + H2O | - |
Cryptococcus sp. S-2 | ? | - |
? | |
| wheat starch + H2O | - |
uncultured bacterium | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 66000, about, recombinant His-tagged chimeric mutant enzyme AmyP-Cr, sequence calculation | uncultured bacterium |
| ? | x * 66000, about, recombinant His-tagged chimeric mutant enzyme AmyP-Cr, sequence calculation | Cryptococcus sp. S-2 |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Amy-1E | - |
uncultured bacterium |
| amy-CS2 | - |
Cryptococcus sp. S-2 |
| AmyP | - |
uncultured bacterium |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
recombinant His-tagged chimeric mutant enzyme AmyP-Cr | Cryptococcus sp. S-2 |
| 40 | - |
wild-type enzyme AmyP and recombinant His-tagged chimeric mutant enzyme AmyP-Cr | uncultured bacterium |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
half-life of chimeric mutant AmyP-Cr at 40°C is 12 h, half-life of wild-type AmyP is 4.5 h | uncultured bacterium |
| 40 | - |
half-life of chimeric mutant AmyP-Cr at 40°C is prolonged by almost 2.6fold when compared with wild-type AmyP from the uncultured bacterium | Cryptococcus sp. S-2 |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
recombinant His-tagged chimeric mutant enzyme AmyP-Cr | Cryptococcus sp. S-2 |
| 7.5 | - |
wild-type enzyme AmyP and recombinant His-tagged chimeric mutant enzyme AmyP-Cr | uncultured bacterium |
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