Recombinant Production of Hyprthermophilic Enzymes: Effects of Temperature and Pressures

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9410687 Robb This project is on research to study the production of enzymes from extremophiles by expression of these enzymes in mesophiles, using recombinant DNA technology. The objective is to make available large quantities of these enzymes for biophysical and biochemical characterization and ultimately for enzymatic applications, without the need for culturing extremophiles directly. The specific objectives of this research include: (1) to clone and identify gene sequences from the hyperthermophile Pyrococcus furiosus and related thermophiles; (2) to express genes in heterologous systems at high pressure, and characterize recombinant enzymes under elevated pressure and temperature to establish optimal specific activity and assembly; and (3) to develop bioprocessing methods for obtaining enzymes in sufficient quantity for structural determination and biotechnological applications. ***