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Literature summary extracted from

  • Marsic, D.; Flaman, J.M.; Ng, J.D.
    New DNA polymerase from the hyperthermophilic marine archaeon Thermococcus thioreducens (2008), Extremophiles, 12, 775-788.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.7 expressed in Escherichia coli BL21(DE3) cells Thermococcus thioreducens

General Stability

EC Number General Stability Organism
2.7.7.7 the enzyme is 20% less active in HEPES and MOPS and 50% less active in phosphate buffer at concentrations of 10 mM, in all cases activity decreases with increased buffer concentration Thermococcus thioreducens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.7 0.00000066
-
 DNAn
-
 Thermococcus thioreducens
2.7.7.7 0.0105
-
 dNTP
-
 Thermococcus thioreducens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.7 K+ over 80% of maximal activity is obtained with KCl concentrations of 50-80 mM, with a maximum at 60 mM Thermococcus thioreducens
2.7.7.7 Mg2+ no activity is detected in the absence of magnesium, activity is maximal between 1.5 and 3.0 mM MgCl2 Thermococcus thioreducens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.7 89980
-
 mature form of PolB, calculated from amino acid sequence Thermococcus thioreducens

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.7 Thermococcus thioreducens A0SXL5 strain ATCC BAA-394, DSM 14981
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.7 HiTrap heparin column chromatography, HiTrap Q column chromatography, and Sephacryl S-200 gel filtration Thermococcus thioreducens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn
-
 Thermococcus thioreducens diphosphate + DNAn+1
-
 ?
2.7.7.7 dNTP + DNAn
-
 Thermococcus thioreducens diphosphate + DNAn+1
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.7 DNA polymerase
-
 Thermococcus thioreducens
2.7.7.7 PolB
-
 Thermococcus thioreducens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.7 72 76 maximal DNA polymerase activity is observed between 72 and 76°C, above which activity decreases abruptly Thermococcus thioreducens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.7.7 95
-
 apparent half-life of 22 h at 95°C with no detectable change during the first 20 h of incubation Thermococcus thioreducens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.7.7 6.5 9 more than 80% of maximal activity is obtained between pH 7.8 and 9.0, at 25°C, which corresponds to pH 6.5-7.7 at 72°C when Tris is used as a buffer Thermococcus thioreducens