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 | 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 | Comment | Organism | Structure |
---|---|---|---|
Ag+ | - |
Geobacillus sp. 4j | |
Cu2+ | - |
Geobacillus sp. 4j | |
Mn2+ | - |
Geobacillus sp. 4j |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Geobacillus sp. 4j | - |
- |
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 | 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 (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 | 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 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 | 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 | 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 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 Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 100 | activity range, profile overview | Geobacillus sp. 4j |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
- |
Geobacillus sp. 4j |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 11 | activity range, profile overview | Geobacillus sp. 4j |