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Literature summary extracted from

  • Nishioka, M.; Mizuguchi, H.; Fujiwara, S.; Komatsubara, S.; Kitabayashi, M.; Uemura, H.; Takagi, M.; Imanaka, T.
    Long and accurate PCR with a mixture of KOD DNA polymerase and its exonuclease deficient mutant enzyme (2001), J. Biotechnol., 88, 141-149.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.7.7.7 molecular biology long and accurate PCR can be achieved with a mixture of wild type DNA polymerase from Thermococcus kodakaraensis and its exonuclease deficient mutant enzyme N210D is utilized (at the ratio of 1:40) Thermococcus kodakarensis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.7
-
 Thermococcus kodakarensis

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.7 N210D the mutant enzyme shows very weak 3'-5'-exonuclease activity compared to the wild-type enzyme (0.1%). No significant difference in DNA polymerase activity as compared with that of the wild-type enzyme. Mutation frequency in PCR becomes higher as 3'–5' exonuclease activity decreases Thermococcus kodakarensis
2.7.7.7 Y311F the mutant enzyme shows very weak 3'-5'-exonuclease activity compared to the wild-type enzyme (0.01%). No significant difference in DNA polymerase activity as compared with that of the wild-type enzyme Thermococcus kodakarensis

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.7 Thermococcus kodakarensis P77933
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.7
-
 Thermococcus kodakarensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.7 deoxynucleoside triphosphate + DNAn the DNA polymerase also shows 3'-5' exonuclease activity. The 3'–5' exonuclease activity of the DNA polymerase is important for DNA elongation with high fidelity Thermococcus kodakarensis diphosphate + DNAn+1
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.7 75
-
 3'-5' exonuclease assay at Thermococcus kodakarensis
2.7.7.7 75
-
 DNA polymerase assay at Thermococcus kodakarensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.7 7.5
-
 DNA polymerase assay at Thermococcus kodakarensis
2.7.7.7 8
-
 3'-5' exonuclease assay at Thermococcus kodakarensis