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Literature summary extracted from

  • Jiang, T.; Cai, M.; Huang, M.; He, H.; Lu, J.; Zhou, X.; Zhang, Y.
    Characterization of a thermostable raw-starch hydrolyzing alpha-amylase from deep-sea thermophile Geobacillus sp. (2015), Protein Expr. Purif., 114, 15-22 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.1 DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic tree, recombinant expression of secereted His6-tagged mutant Gs4j-amyA N-terminally fused to OmpA signal peptide in Escherichia coli strain BL21(DE3) yielding 22fold increased enzyme production and activity as compared to the wild-type strain Geobacillus sp. 4j

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.1 additional information an N-terminally truncated form of Geobacillus sp. 4j alpha-amylase (Gs4j-amyA) is fused at its N-terminal end with the signal peptide of outer membrane protein A (OmpA) of Escherichia coli Geobacillus sp. 4j

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.1 Ag+
-
 Geobacillus sp. 4j
3.2.1.1 Cu2+
-
 Geobacillus sp. 4j
3.2.1.1 Mn2+
-
 Geobacillus sp. 4j

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.1 extracellular
-
 Geobacillus sp. 4j
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.1 Ca2+ Ca2+ at 10 mM has only a slightly stimulating effect, the enzyme is Ca2+-independent Geobacillus sp. 4j

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.1 Geobacillus sp. 4j
-
 first isolated from the sub-seafloor sediments, which have been collected from WP0104 site at the western Pacific warm pool (8x0253'08''N, 142x0259'51''E, 2068 m of water depth)
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.1 recombinant extracellular His-tagged mutant enzyme Gs4j-amyA from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation and nickel affinity chromatography Geobacillus sp. 4j

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.1 additional information strain 4j is grown in an alpha-amylase producing medium containing soluble starch as the sole carbon source at 60°C Geobacillus sp. 4j
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.1 8600
-
 purified recombinant mutant enzyme Gs4j-amyA, pH 5.5, 60°C Geobacillus sp. 4j

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.1 amylopectin + H2O 67.9% activity compared to gelatinized soluble starch Geobacillus sp. 4j ?
-
 ?
3.2.1.1 amylose + H2O 45.5% activity compared to gelatinized soluble starch Geobacillus sp. 4j ?
-
 ?
3.2.1.1 cassava raw starch + H2O
-
 Geobacillus sp. 4j ?
-
 ?
3.2.1.1 corn raw starch + H2O
-
 Geobacillus sp. 4j ?
-
 ?
3.2.1.1 gelatinized soluble starch + H2O best substrate Geobacillus sp. 4j ?
-
 ?
3.2.1.1 additional information no activity with pullulan and sucrose Geobacillus sp. 4j ?
-
 ?
3.2.1.1 pea raw starch + H2O
-
 Geobacillus sp. 4j ?
-
 ?
3.2.1.1 potato raw starch + H2O
-
 Geobacillus sp. 4j ?
-
 ?
3.2.1.1 raw soluble starch + H2O 22% activity compared to gelatinized soluble starch Geobacillus sp. 4j ?
-
 ?
3.2.1.1 sweet potato raw starch + H2O
-
 Geobacillus sp. 4j ?
-
 ?
3.2.1.1 wheat raw starch + H2O
-
 Geobacillus sp. 4j ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.1 ? x * 62000, recombinant His6-tagged mutant Gs4j-amyA N-terminally fused to OmpA signal peptide, SDS-PAGE, x * 58770, sequence calculation Geobacillus sp. 4j

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.1 60 80 gelatinized soluble starch is the best substrate at 60°C. The enzyme is optimally active at 70°C, when acting on amylopectin and raw potato starch. As for hydrolysis of corn, cassava, sweet potato, wheat, pea starches and amylose, the optimum reaction temperature is 80°C Geobacillus sp. 4j

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.1 30 100 activity range, profile overview Geobacillus sp. 4j

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.1 80
-
 purified recombinant His6-tagged mutant Gs4j-amyA N-terminally fused to OmpA signal peptide, half-life (t1/2) is 4.25 h Geobacillus sp. 4j

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.1 5.5
-
-
 Geobacillus sp. 4j

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.1 4 11 activity range, profile overview Geobacillus sp. 4j