Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.7.7.7 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)

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

General Stability

General Stability Organism
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]

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

Metals/Ions

Metals/Ions Comment Organism Structure
K+ over 80% of maximal activity is obtained with KCl concentrations of 50-80 mM, with a maximum at 60 mM Thermococcus thioreducens
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]

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

Organism

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

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
DNA polymerase
-
Thermococcus thioreducens
PolB
-
Thermococcus thioreducens

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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

pH Minimum pH Maximum Comment Organism
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