Activating Compound | Comment | Organism | Structure |
---|---|---|---|
AMP | slight activation | Acetivibrio thermocellus | |
DTT | activates | Acetivibrio thermocellus |
Cloned (Comment) | Organism |
---|---|
ORF Cthe0357, DNA and amino acid sequence determination and analysis, expression of C-terminally His-tagged enzyme in Escherichia coli, expression of cellulose-binding module-intein-alphaGP fusion protein | Acetivibrio thermocellus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | slight activation | Acetivibrio thermocellus | |
Mn2+ | strong inhibition at 1 mM | Acetivibrio thermocellus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.054 | - |
alpha-D-glucose 1-phosphate | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
0.3 | - |
starch | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
0.39 | - |
dextrin DP19 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
0.73 | - |
dextrin DP14 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
1.2 | - |
dextrin DP14 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
1.5 | - |
starch | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
1.8 | - |
dextrin DP4 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
1.9 | - |
dextrin DP19 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
7.7 | - |
phosphate | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
17.1 | - |
dextrin DP4 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | activates at 1 mM | Acetivibrio thermocellus | |
additional information | the enzyme is not affected by 1 mM of Ca2+, Cu2+, Zn2+, and Ni2+ | Acetivibrio thermocellus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
[(1->4)-alpha-D-glucosyl]n + phosphate | Acetivibrio thermocellus | - |
[(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate | - |
r | |
[(1->4)-alpha-D-glucosyl]n + phosphate | Acetivibrio thermocellus ATCC 27405 | - |
[(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acetivibrio thermocellus | - |
ORF Cthe0357 | - |
Acetivibrio thermocellus ATCC 27405 | - |
ORF Cthe0357 | - |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His-tagged enzyme from Escherichia coli by nickel affinity chromatography, recombinant cellulose-binding moduleintein-alphaGP fusion protein by affinity adsorption on amorphous cellulose followed by intein self-cleavage | Acetivibrio thermocellus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.08 | - |
purified recombinant detagged enzyme, pH 6.8, 50°C | Acetivibrio thermocellus |
2.22 | - |
purified recombinant His-tagged enzyme, pH 6.8, 50°C | Acetivibrio thermocellus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dextrin DP13 + alpha-D-glucose 1-phosphate | prolongation of the dextrin molecule with one glucose monomer | Acetivibrio thermocellus | dextrin DP14 + phosphate | - |
r | |
dextrin DP13 + alpha-D-glucose 1-phosphate | prolongation of the dextrin molecule with one glucose monomer | Acetivibrio thermocellus ATCC 27405 | dextrin DP14 + phosphate | - |
r | |
dextrin DP14 + phosphate | - |
Acetivibrio thermocellus | dextrin DP13 + alpha-D-glucose 1-phosphate | - |
r | |
dextrin DP14 + phosphate | - |
Acetivibrio thermocellus ATCC 27405 | dextrin DP13 + alpha-D-glucose 1-phosphate | - |
r | |
dextrin DP18 + alpha-D-glucose 1-phosphate | prolongation of the dextrin molecule with one glucose monomer | Acetivibrio thermocellus | dextrin DP19 + phosphate | - |
r | |
dextrin DP19 + phosphate | - |
Acetivibrio thermocellus | dextrin DP18 + alpha-D-glucose 1-phosphate | - |
r | |
dextrin DP3 + alpha-D-glucose 1-phosphate | prolongation of the dextrin molecule with one glucose monomer | Acetivibrio thermocellus | dextrin DP4 + phosphate | - |
r | |
dextrin DP4 + phosphate | - |
Acetivibrio thermocellus | dextrin DP3 + alpha-D-glucose 1-phosphate | - |
r | |
additional information | broad substrate specificities for different chain length dextrins and soluble starch, enzyme efficiency on dextrin DP4 is approximately 5fold lower than that on dextrin DP14 although kcat on dextrin DP4 is higher than on dextrin DP14 | Acetivibrio thermocellus | ? | - |
? | |
additional information | broad substrate specificities for different chain length dextrins and soluble starch, enzyme efficiency on dextrin DP4 is approximately 5fold lower than that on dextrin DP14 although kcat on dextrin DP4 is higher than on dextrin DP14 | Acetivibrio thermocellus ATCC 27405 | ? | - |
? | |
starch + alpha-D-glucose 1-phosphate | prolongation of the starch molecule with one glucose monomer | Acetivibrio thermocellus | starch + phosphate | - |
r | |
starch + alpha-D-glucose 1-phosphate | prolongation of the starch molecule with one glucose monomer | Acetivibrio thermocellus ATCC 27405 | starch + phosphate | - |
r | |
[(1->4)-alpha-D-glucosyl]n + phosphate | - |
Acetivibrio thermocellus | [(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate | - |
r | |
[(1->4)-alpha-D-glucosyl]n + phosphate | - |
Acetivibrio thermocellus ATCC 27405 | [(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
alpha-glucan phosphorylase | - |
Acetivibrio thermocellus |
alphaGP | - |
Acetivibrio thermocellus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
synthesis direction | Acetivibrio thermocellus |
80 | - |
degradation direction | Acetivibrio thermocellus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 90 | activity range | Acetivibrio thermocellus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
purified recombinant enzyme, loss of 10% activity within 120 min, stable | Acetivibrio thermocellus |
50 | - |
half-life without additives is about 2 h, and is extended by 45fold to 90 h through a combined addition of 0.1 mM Mg2+, 5 mM DTT, 1% NaCl, 0.1% Triton X-100, and 1 mg/mL BSA | Acetivibrio thermocellus |
70 | - |
purified recombinant enzyme, half-ife is about 40 min, inactivation after 120 min | Acetivibrio thermocellus |
80 | - |
purified recombinant enzyme, loss of over 90% activity within 10 min, inactivation after 60 min | Acetivibrio thermocellus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.1 | - |
starch | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
3.6 | - |
dextrin DP14 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
4.2 | - |
alpha-D-glucose 1-phosphate | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
6.3 | - |
starch | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
6.6 | - |
dextrin DP19 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
8.1 | - |
dextrin DP19 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
8.2 | - |
phosphate | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
8.4 | - |
dextrin DP4 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
9.3 | - |
dextrin DP14 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
11 | - |
dextrin DP4 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 6.5 | synthesis direction | Acetivibrio thermocellus |
7 | 7.5 | degradation direction | Acetivibrio thermocellus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 10 | activity range | Acetivibrio thermocellus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.64 | - |
dextrin DP4 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
1.1 | - |
phosphate | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
1.5 | - |
starch | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
3 | - |
dextrin DP14 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
3.5 | - |
dextrin DP19 | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
4.7 | - |
dextrin DP4 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
7.7 | - |
alpha-D-glucose 1-phosphate | pH 6.8, 50°C, synthesis reaction | Acetivibrio thermocellus | |
13 | - |
dextrin DP14 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
21 | - |
starch | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus | |
21 | - |
dextrin DP19 | pH 7.2, 50°C, degradation reaction | Acetivibrio thermocellus |