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Literature summary for 2.7.7.7 extracted from

  • Pavlov, A.R.; Pavlova, N.V.; Kozyavkin, S.A.; Slesarev, A.I.
    Cooperation between catalytic and DNA binding domains enhances thermostability and supports DNA synthesis at higher temperatures by thermostable DNA polymerases (2012), Biochemistry, 51, 2032-2043.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of the N-terminal part of the LF Bst polymerase gene in Escherichia coli strain BL21 Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
additional information engineered DNA polymerases are obtained by the flexible attachment of helix-hairpin-helix DNA binding domains of topoisomerase V of Methanopyrus kandleri to catalytic domains of DNA polymerases, domain fusion using the Taq and Pfu DNA polymerases or the phi29 DNA polymerase. The chimeric polymerases retain high processivity at high concentrations of salts and other inhibitors of DNA synthesis, such as phenol, blood, and DNA intercalating dyes, domain attachment has a potential to greatly increase the thermostability of chimeric DNA polymerases Methanopyrus kandleri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
deoxynucleoside triphosphate + DNAn Geobacillus stearothermophilus
-
diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn Methanopyrus kandleri
-
diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn Geobacillus stearothermophilus Donc
-
diphosphate + DNAn+1
-
?

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
ATCC12980D-5
-
Geobacillus stearothermophilus Donc
-
ATCC12980D-5
-
Methanopyrus kandleri
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminal part of the LF Bst polymerase from Escherichia coli strain BL21 by anion exchange chromatography Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
deoxynucleoside triphosphate + DNAn
-
Geobacillus stearothermophilus diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn
-
Methanopyrus kandleri diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn standard substrate PTJ1 and substrate PTJ2 Geobacillus stearothermophilus diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn standard substrate PTJ1 and substrate PTJ2 Methanopyrus kandleri diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn
-
Geobacillus stearothermophilus Donc diphosphate + DNAn+1
-
?
deoxynucleoside triphosphate + DNAn standard substrate PTJ1 and substrate PTJ2 Geobacillus stearothermophilus Donc diphosphate + DNAn+1
-
?

Synonyms

Synonyms Comment Organism
Bst DNA polymerase
-
Geobacillus stearothermophilus
Mka polB
-
Methanopyrus kandleri

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Geobacillus stearothermophilus
25
-
assay at Methanopyrus kandleri

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 95 thermostability and half-life of the LF Bst polymerase with the attached TopoV's HhH C2 domain increases almost 8fold, as compared to the Bst LF polymerase alone. The effect can be seen up to 95°C. The stabilizing effect of the C3 domain in N-TopoTaq completely disappears if K-Glu is replaced by 0.5 M NaCl, and the stability of N-TopoTaq does not differ from that of the catalytic domain alone, addition of 1 M betaine restores the stabilization provided by the C3 domain. Electrostatic enhancement of diffusion-controlled association. Thermal stabilization of chimeric polymerases occurs due to charge-independant interactions of the polymerase domain and HhH domains Geobacillus stearothermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Geobacillus stearothermophilus
8
-
assay at Methanopyrus kandleri

General Information

General Information Comment Organism
evolution the enzyme belongs to the DNA polymerase family B Methanopyrus kandleri