Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain ORIGAMI(DE3)pLysS | Ferroplasma acidiphilum |
Protein Variants | Comment | Organism |
---|---|---|
A576P | the mutation hardly affects ligation activity at pH 3.0 | Ferroplasma acidiphilum |
D162E | the mutation hardly affects ligation activity at pH 3.0 | Ferroplasma acidiphilum |
E134L | the mutation causes a 60% reduction at pH 3.0, with no net change in iron content and purple color, the mutant has an activity optimum of pH 5.0 and a 1.5fold higher turnover number as the wild type enzyme | Ferroplasma acidiphilum |
N255G | the mutation hardly affects ligation activity at pH 3.0 | Ferroplasma acidiphilum |
T491S | the mutation hardly affects ligation activity at pH 3.0 | Ferroplasma acidiphilum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe3+ | contains two Fe3+-tyrosinate centers, reduction of the Fe3+ to Fe2+ results in an 80% decrease in DNA substrate binding and an increase in the pH activity optimum to 5.0 | Ferroplasma acidiphilum | |
additional information | catalytic activity neither depends on nor is stimulated by added Mg2+ or K+ | Ferroplasma acidiphilum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ferroplasma acidiphilum | Q2PCE4 | - |
- |
Ferroplasma acidiphilum YT / DSM 12658 | Q2PCE4 | - |
- |
Purification (Comment) | Organism |
---|---|
Resource Q column chromatography and Superdex-200 gel filtration | Ferroplasma acidiphilum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | - |
Ferroplasma acidiphilum | AMP + diphosphate + (deoxyribonucleotide)m+n | - |
? | |
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | - |
Ferroplasma acidiphilum YT / DSM 12658 | AMP + diphosphate + (deoxyribonucleotide)m+n | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DNA ligase | - |
Ferroplasma acidiphilum |
Lig | - |
Ferroplasma acidiphilum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.025 | - |
(deoxyribonucleotide)n | mutant enzyme E143L | Ferroplasma acidiphilum | |
0.048 | - |
(deoxyribonucleotide)n | mutant enzyme T491S | Ferroplasma acidiphilum | |
0.05 | - |
(deoxyribonucleotide)n | mutant enzyme D162E | Ferroplasma acidiphilum | |
0.051 | - |
(deoxyribonucleotide)n | mutant enzyme A576P | Ferroplasma acidiphilum | |
0.051 | - |
(deoxyribonucleotide)n | mutant enzyme N255G | Ferroplasma acidiphilum | |
0.056 | - |
(deoxyribonucleotide)n | wild type enzyme | Ferroplasma acidiphilum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
1.5 | 3 | pH-optimum in vitro | Ferroplasma acidiphilum |
5 | - |
reduction of the Fe3+ to Fe2+ results in an 80% decrease in DNA substrate binding and an increase in the pH activity optimum to 5.0 | Ferroplasma acidiphilum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
1.5 | 5 | ligation activities, at 40°C under conditions of substrate saturation, are highest at pH 2.5-3.0, only slightly lower at pH 1.5-2.0 (about 80%) and are practically undetectable above pH 5.0 | Ferroplasma acidiphilum |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
2 | 3 | LigFa is acid-tolerant and retains about 95% of its activity after incubation at pH 2.0-3.0 for 10 h but is unstable at higher pH levels | Ferroplasma acidiphilum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | ATP is preferred over NAD+ | Ferroplasma acidiphilum | |
NAD+ | uses also NAD+ as cofactor although ATP is preferred | Ferroplasma acidiphilum |