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Overall, 30 strains of hyperthermophilic archaea, representing seven species of the genera Thermococcus, Desulfurococcus, Thermoproteus, and Acidilobus, were tested for the presence of thermostable DNA polymerases. Thermostabilities of the polymerases varied distinctly among the strains within one species. Polymerases of five strains retained 60–100% activity upon incubation of the preparations at 95°C for 120 min. A new DNA polymerase was isolated from the strain Thermococcus litoralis Sh1AM, possessing the enzyme with the most promising properties, and characterized. Molecular weight of the enzyme is 90–100% kDa. The purified DNA polymerase preserved 50% of the initial activity upon incubation at 95°C for 120 min. The polymerase isolated displayed an associated 3’–5’ exonuclease activity. The error rate when extending a DNA strand was at least twofold lower compared with Taq polymerase. The main physicochemical and enzymatic properties of the new polymerase are similar to the known DNA polymerases of family B.
Applied Biochemistry and Microbiology – Springer Journals
Published: Feb 16, 2005
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