Cloned (Comment) | Organism |
---|---|
pol beta in vitro/in vivo dual expression using plasmid pIVEX, overexpression in Escherichia coli strain BL21(DE3), strain BL21(DE3)pLysS, and strain KRX, and in vitro as a C-terminal His-tagged protein. Pol beta expressed in vitro is subject to proteolysis, therefore, bacterial overexpression is used to produce the protein for kinetic analyses. KRX cells are preferred because of their reduced propensity for leaky expression of pol beta | Danio rerio |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00132 | - |
dTTP | pH 8.8, 30°C, recombinant enzyme, in presence of Mn2+ | Danio rerio | |
0.01469 | - |
dTTP | pH 8.8, 30°C, recombinant enzyme, in presence of Mg2+ | Danio rerio |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
KCl | activates best at 10 mM | Danio rerio | |
Mg2+ | activates, Mn2+ shows a 8.6fold higher catalytic efficiency than Mg2+ | Danio rerio | |
Mn2+ | activates, Mn2+ shows a 8.6fold higher catalytic efficiency than Mg2+ | Danio rerio | |
NaCl | activates best at 50 mM | Danio rerio |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
deoxynucleoside triphosphate + DNAn | Danio rerio | - |
diphosphate + DNAn+1 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Danio rerio | Q6DRD3 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
deoxynucleoside triphosphate + DNAn | - |
Danio rerio | diphosphate + DNAn+1 | - |
? | |
dTTP + DNAn | - |
Danio rerio | diphosphate + DNAn+1 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DNA polymerase beta | - |
Danio rerio |
pol beta | - |
Danio rerio |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Danio rerio |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0096 | - |
dTTP | pH 8.8, 30°C, recombinant enzyme, in presence of Mn2+ | Danio rerio | |
0.0123 | - |
dTTP | pH 8.8, 30°C, recombinant enzyme, in presence of Mg2+ | Danio rerio |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Danio rerio |
General Information | Comment | Organism |
---|---|---|
evolution | compared to mammalian pol betas, the Danio rerio enzyme contains a P63D amino acid substitution. This substitution lies in a hairpin sequence within an 8-kDa domain, likely to be important in DNA binding | Danio rerio |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.83 | - |
dTTP | pH 8.8, 30°C, recombinant enzyme, in presence of Mg2+ | Danio rerio | |
7.16 | - |
dTTP | pH 8.8, 30°C, recombinant enzyme, in presence of Mn2+ | Danio rerio |