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Literature summary for 3.2.1.20 extracted from
- A new GH13 alpha-glucosidase from alkaliphilic Bacillus pseudofirmus 703 with both exo-alpha-1, 4-glucosidase and oligo-1, 6-glucosidase activities toward amylopectin (2017), Int. J. Biol. Macromol., 101, 973-982 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene amy112, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Alkalihalophilus pseudofirmus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E257A | site-directed mutagenesis, mutation of a catalytic residue, inactive mutant | Alkalihalophilus pseudofirmus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Cd2+ | strong inhibition at 5 mM | Alkalihalophilus pseudofirmus |  |
| Co2+ | weak inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
| Cu2+ | almost complete inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
| DMSO | weak inhibition at 5% | Alkalihalophilus pseudofirmus | |
| EDTA | moderate inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
| ethanol | moderate inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
| glycerol | moderate inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
| Hg2+ | almost complete inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
| Isopropyl alcohol | strong inhibition at 5% | Alkalihalophilus pseudofirmus | |
| methanol | moderate inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
| SDS | strong inhibition at 0.0025% | Alkalihalophilus pseudofirmus | |
| Zn2+ | moderate inhibition at 5 mM | Alkalihalophilus pseudofirmus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Lineweaver-Burk kinetics | Alkalihalophilus pseudofirmus | |
| 2.99 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 7.0, 40°C | Alkalihalophilus pseudofirmus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| K+ | addition of K+ ions improves the amy112 activity by 12% | Alkalihalophilus pseudofirmus | |
| additional information | Li+, Ca2+, and Mg2+ ions show no significant effect on enzyme activity | Alkalihalophilus pseudofirmus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Alkalihalophilus pseudofirmus | A0A1S6JYL1 | - |
- |
| Alkalihalophilus pseudofirmus 703 | A0A1S6JYL1 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type enzyme 5.72fold and of mutant enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, desalting gel filtration, and anion exchange chromatography, to homogeneity | Alkalihalophilus pseudofirmus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl alpha-D-glucopyranoside + H2O | - |
Alkalihalophilus pseudofirmus | 4-nitrophenol + D-glucose | - |
? | |
| 4-nitrophenyl alpha-D-glucopyranoside + H2O | - |
Alkalihalophilus pseudofirmus 703 | 4-nitrophenol + D-glucose | - |
? | |
| amylopectin + H2O | from maize, best substrate | Alkalihalophilus pseudofirmus | D-glucose + ? | - |
? | |
| amylopectin + H2O | from maize, best substrate | Alkalihalophilus pseudofirmus 703 | D-glucose + ? | - |
? | |
| maltose + H2O | - |
Alkalihalophilus pseudofirmus | 2 D-glucose | - |
? | |
| maltose + H2O | - |
Alkalihalophilus pseudofirmus 703 | 2 D-glucose | - |
? | |
| maltotriose + H2O | - |
Alkalihalophilus pseudofirmus | maltose + D-glucose | - |
? | |
| additional information | the alpha-glucosidase (EC 3.2.1.20) from Bacillus pseudofirmus strain 703 also shows oligo-alpha-1,6-glucosidase activity (EC 3.2.1.10), having both exo-alpha-1,4-glucosidase and oligo-l,6-glucosidase activities, but it shows no transglycosylation activity. The enzyme exhibits high activity against amylopectin from maize and soluble starch, but no activity against amylose from potato, pullulan, carboxymethylcellulose, and beachwood xylan. Amy112 hydrolyzes alpha-1,6-glucosidic linkages less effective than alpha-1,4-glucosidic linkages. Hydrolysates of amylopectin from maize, maltose, and maltotriose are mainly glucose | Alkalihalophilus pseudofirmus | ? | - |
? | |
| additional information | the alpha-glucosidase (EC 3.2.1.20) from Bacillus pseudofirmus strain 703 also shows oligo-alpha-1,6-glucosidase activity (EC 3.2.1.10), having both exo-alpha-1,4-glucosidase and oligo-l,6-glucosidase activities, but it shows no transglycosylation activity. The enzyme exhibits high activity against amylopectin from maize and soluble starch, but no activity against amylose from potato, pullulan, carboxymethylcellulose, and beachwood xylan. Amy112 hydrolyzes alpha-1,6-glucosidic linkages less effective than alpha-1,4-glucosidic linkages. Hydrolysates of amylopectin from maize, maltose, and maltotriose are mainly glucose | Alkalihalophilus pseudofirmus 703 | ? | - |
? | |
| soluble starch + H2O | 76.2% activity compared to amylopectin | Alkalihalophilus pseudofirmus | ? | - |
? | |
| soluble starch + H2O | 76.2% activity compared to amylopectin | Alkalihalophilus pseudofirmus 703 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Amy112 | - |
Alkalihalophilus pseudofirmus |
| exo-alpha-l, 4-glucosidase | - |
Alkalihalophilus pseudofirmus |
| exo-alpha-l,4-glucosidase | - |
Alkalihalophilus pseudofirmus |
| More | cf. EC 3.2.1.10 | Alkalihalophilus pseudofirmus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 40 | - |
Alkalihalophilus pseudofirmus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 50 | profile overview, 50% of maximal activity at 35-45°C, maximal activity at 30-40°C | Alkalihalophilus pseudofirmus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 50 | Amy112 retains most of its activity after 30 min pre-incubation at 30°C and 40°C, but loses activity rapidly after 30 min above 50°C | Alkalihalophilus pseudofirmus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Alkalihalophilus pseudofirmus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6.2 | 8.6 | over 50% of maximal activity within this range, profile overview | Alkalihalophilus pseudofirmus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the glycoside hydrolase family 13, GH13 | Alkalihalophilus pseudofirmus |
| additional information | enzyme three-dimensional structure, structure homology modeling based on GSJ from Geobacillus sp. HTA-462 (PDB ID 2ZE0), the predicted catalytic triad is comprised by Asp200, Glu257, and Asp327 in Amy112 | Alkalihalophilus pseudofirmus |
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