EC Number | Cloned (Comment) | Organism |
---|---|---|
3.2.1.135 | Escherichia coli NovaBlue and Rosetta (DE3) pLysS, cloning and expression host, respectively, with plasmid pET-20b(+), in LB broth, 37°C | Thermoanaerobacter ethanolicus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.2.1.135 | additional information | R855 C-terminal truncated (100 amino acids) mutant R855 of full length TetApuM955, produced with trypsin-like proteolytic cleavage reaction | Thermoanaerobacter ethanolicus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.135 | additional information | - |
pullulan | 11.6 mg/ml, TetApuM955 | Thermoanaerobacter ethanolicus | |
3.2.1.135 | additional information | - |
pullulan | 16.0 mg/ml, TetApuR855 | Thermoanaerobacter ethanolicus | |
3.2.1.135 | additional information | - |
starch | 5.2 mg/ml, TetApuM955 | Thermoanaerobacter ethanolicus | |
3.2.1.135 | additional information | - |
starch | 6.0 mg/ml, TetApuR855 | Thermoanaerobacter ethanolicus |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.2.1.135 | 85000 | - |
theoretically calculated mass of truncated TetApuM855, corroborated by SDS-PAGE | Thermoanaerobacter ethanolicus |
3.2.1.135 | 107800 | - |
calculated mass of matured full length enzmye TetAbuM955, corroborated by SDS-PAGE, 13 N-terminal amino acids (signal peptide) cleaved off proteolytically, DNA fragment 2882 bp | Thermoanaerobacter ethanolicus |
3.2.1.135 | 107818 | - |
x * 107818, SDS-PAGE | Thermoanaerobacter ethanolicus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.2.1.135 | Thermoanaerobacter ethanolicus | - |
TetApuM955 gene of 2.9 kb | - |
3.2.1.135 | Thermoanaerobacter ethanolicus 39E | - |
TetApuM955 gene of 2.9 kb | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.2.1.135 | BD TALON spin column for 6x His-tag protein affinity purification, simultaneous SDS-PAGE zymogram with 1% soluble starch | Thermoanaerobacter ethanolicus |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.2.1.135 | additional information | - |
117400000000 micromol/min/mol enzyme, TetApuR855, soluble starch | Thermoanaerobacter ethanolicus |
3.2.1.135 | additional information | - |
11750000000 micromol/min/mol enzyme, TetApuR855, raw starch | Thermoanaerobacter ethanolicus |
3.2.1.135 | additional information | - |
11860000000 micromol/min/mol enzyme, TetApuM955, raw starch | Thermoanaerobacter ethanolicus |
3.2.1.135 | additional information | - |
119400000000 micromol/min/mol enzyme, TetApuM955, soluble starch | Thermoanaerobacter ethanolicus |
3.2.1.135 | additional information | - |
234200000000 micromol/min/mol enzyme, TetApuM955, pullulan | Thermoanaerobacter ethanolicus |
3.2.1.135 | additional information | - |
243000000000 micromol/min/mol enzyme, TetApuR855, pullulan | Thermoanaerobacter ethanolicus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.1.135 | glycogen + H2O | 50 mM sodium acetate buffer, pH 6.0, 5 mM CaCl2, 80°C | Thermoanaerobacter ethanolicus | maltooligosaccharides | - |
? | |
3.2.1.135 | glycogen + H2O | 50 mM sodium acetate buffer, pH 6.0, 5 mM CaCl2, 80°C | Thermoanaerobacter ethanolicus 39E | maltooligosaccharides | - |
? | |
3.2.1.135 | pullulan + H2O | 50 mM sodium acetate buffer, pH 6.0, 5 mM CaCl2, 75°C | Thermoanaerobacter ethanolicus | maltooligosaccharides | - |
? | |
3.2.1.135 | pullulan + H2O | 50 mM sodium acetate buffer, pH 6.0, 5 mM CaCl2, 75°C | Thermoanaerobacter ethanolicus 39E | maltooligosaccharides | - |
? | |
3.2.1.135 | soluble and raw starch + H2O | 50 mM sodium acetate buffer, pH 6.0, 5 mM CaCl2, 80°C | Thermoanaerobacter ethanolicus | maltose + maltotriose + maltotetraose + maltopentaose + maltohexaose + maltoheptaose + ? | - |
? | |
3.2.1.135 | soluble and raw starch + H2O | 50 mM sodium acetate buffer, pH 6.0, 5 mM CaCl2, 80°C | Thermoanaerobacter ethanolicus 39E | maltose + maltotriose + maltotetraose + maltopentaose + maltohexaose + maltoheptaose + ? | - |
? | |
3.2.1.135 | starch + H2O | - |
Thermoanaerobacter ethanolicus | maltose + ? | - |
? | |
3.2.1.135 | starch + H2O | - |
Thermoanaerobacter ethanolicus 39E | maltose + ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.2.1.135 | ? | x * 85000, SDS-PAGE | Thermoanaerobacter ethanolicus |
3.2.1.135 | ? | x * 107818, SDS-PAGE | Thermoanaerobacter ethanolicus |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.2.1.135 | amylopullulanase | - |
Thermoanaerobacter ethanolicus |
3.2.1.135 | TetApuM955 | full length enzyme | Thermoanaerobacter ethanolicus |
3.2.1.135 | TetApuR855 | truncated enzyme | Thermoanaerobacter ethanolicus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.2.1.135 | 75 | - |
75-80°C optimal for full length and truncated enzyme | Thermoanaerobacter ethanolicus |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.2.1.135 | 80 | - |
no temperature sensitivity difference between truncated and full length enzyme below and at 80°C | Thermoanaerobacter ethanolicus |
3.2.1.135 | 88 | - |
onset of denaturation of full length TetApuM955 | Thermoanaerobacter ethanolicus |
3.2.1.135 | 90 | - |
onset of denaturation of truncated TetApuR885, it is more thermostable than full length TetApuM955 above 90°C | Thermoanaerobacter ethanolicus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.135 | 298.3 | - |
starch | TetApuM955 | Thermoanaerobacter ethanolicus | |
3.2.1.135 | 313.3 | - |
starch | TetApuR855 | Thermoanaerobacter ethanolicus | |
3.2.1.135 | 828.3 | - |
pullulan | TetApuM955 | Thermoanaerobacter ethanolicus | |
3.2.1.135 | 1010 | - |
pullulan | TetApuR855 | Thermoanaerobacter ethanolicus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.2.1.135 | 6 | - |
full length and truncated enzyme | Thermoanaerobacter ethanolicus |