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Literature summary for 3.2.1.1 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
Search for more literature for 3.2.1.1

Cloned(Commentary)

Cloned (Comment) Organism
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

Protein Variants Comment Organism
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

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

Localization

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

Metals/Ions

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

Organism

Organism UniProt Comment Textmining
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)

Purification (Comment) Organism
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

Source Tissue Comment Organism Textmining
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]

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

Substrates and Products (Substrate)

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

Subunits

Subunits Comment Organism
? 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]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
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]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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

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

pH Range

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