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Literature summary extracted from

  • Ishido, T.; Yamazaki, N.; Ishikawa, M.; Hirano, K.
    Characterization of DNA polymerase beta from Danio rerio by overexpression in E. coli using the in vivo/in vitro compatible pIVEX plasmid (2011), Microb. Cell Fact., 10, 84.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.7 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.7 0.00132
-
 dTTP pH 8.8, 30°C, recombinant enzyme, in presence of Mn2+ Danio rerio
2.7.7.7 0.01469
-
 dTTP pH 8.8, 30°C, recombinant enzyme, in presence of Mg2+ Danio rerio

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.7 KCl activates best at 10 mM Danio rerio
2.7.7.7 Mg2+ activates, Mn2+ shows a 8.6fold higher catalytic efficiency than Mg2+ Danio rerio
2.7.7.7 Mn2+ activates, Mn2+ shows a 8.6fold higher catalytic efficiency than Mg2+ Danio rerio
2.7.7.7 NaCl activates best at 50 mM Danio rerio

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn Danio rerio
-
 diphosphate + DNAn+1
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.7 Danio rerio Q6DRD3
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Danio rerio diphosphate + DNAn+1
-
 ?
2.7.7.7 dTTP + DNAn
-
 Danio rerio diphosphate + DNAn+1
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.7 DNA polymerase beta
-
 Danio rerio
2.7.7.7 pol beta
-
 Danio rerio

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.7 30
-
-
 Danio rerio

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.7 0.0096
-
 dTTP pH 8.8, 30°C, recombinant enzyme, in presence of Mn2+ Danio rerio
2.7.7.7 0.0123
-
 dTTP pH 8.8, 30°C, recombinant enzyme, in presence of Mg2+ Danio rerio

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.7 9
-
-
 Danio rerio

General Information

EC Number General Information Comment Organism
2.7.7.7 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.7.7 0.83
-
 dTTP pH 8.8, 30°C, recombinant enzyme, in presence of Mg2+ Danio rerio
2.7.7.7 7.16
-
 dTTP pH 8.8, 30°C, recombinant enzyme, in presence of Mn2+ Danio rerio