Abstract
Protein Biotechnology The polymerase chain reaction (PCR) is a key tool in medical and biological research. The most common PCR reaction relies on the thermal cycling method that consists of repeated cycles of heating and cooling steps for DNA melting and extension by the DNA polymerase, respectively. The introduction of new DNA polymerases to the market is a major area of development that tremendously helped in improving the performance and quality of PCR. Nonetheless, PCR still requires optimization of salt and metal ion concentrations leaving a room in the market for introducing new DNA polymerases that are robuster in their salt and metal ion concentration dependence. In this study, we will present the characterization of a novel archaeal DNA polymerase from the Red Sea brine-pool (termed BR3) and demonstrate how its enzymatic activity reflects on every aspects of the environment of the brine-pool – high tolerance to concentrations and types of salts and metal ions including utilization of Zn21ions in its active site. These results suggest that the brine-pool microorganisms are likely to contain novel chemical pathways to deal with its exterior harsh conditions. We will further show the mechanism of BR3 polymerase how it was adjusted to be active in harsh condition.