This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Extreme environments have been of considerable interest in recent years due to the discovery that these "uninhabitable" environments are colonized by a diverse community of microorganisms. These microorganisms have been shown to provide valuable extremozymes, enzymes adapted to catalyze reactions under extreme environmental conditions. One well-known extremozyme is Taq polymerase from the hyperthermophile Thermus aquaticus. This research focuses on the little studied, hypersaline, alkaline lakes in western Nebraska. We will isolate and identify potential microorganisms capable of producing extremozymes of interest for numerous biotechnology applications. We will look for esterases, xylanases, cellulases, proteases, and lipases that are active at high pH's. Microorganisms capable of producing these enzymes will be isolated, the enzyme activity characterized, the enzyme purified, and the enzyme gene sequences identified and cloned into Escherichia coli.