Thermostable Esterases and Lipases

We will develop a collection of thermostable esterases and lipases which can be used in chiralsynthesis of new and safer anticancer and antiviral pharmaceuticals. Phase I feasibility will bedemonstrated with discovery of several new enzymes from a variety of extreme- and hyperthermopholicorganisms. These enzymes will be grouped by substrates specificities and kinetic properties. A subsetof enzymes will be characterized further by gene cloning, protein purification, analysis of thermostability,determination of optimum pH and ionic conditions, and preliminary determination of substratespecificities. Their commercial importance will be assessed, and samples will be tested by industrialpharmaceutical research groups. Phase II will involve expansion of the collection for more specificitiesand improved properties. Useful enzymes will be more completely characterized by nucleotidesequencing. Organic solvent profiles, and kinetic characterization including product inhibition. Keyenzymes will be immobilized and adapted into enzyme screening systems and prototype bioreactors totest for stability and reusability. Finally, to further extend applications, genetic selections will beattempted to modify their substrate specificities and kinetic properties. Phase III will involve thecommercialization of the enzymatic activities and adaption and production of the enzymes for large scalechiral chemical synthesis processes.