Phase I of this project was aimed at the development of an extremely high throughout screening system based on Fluorescence activated cell sorting for the discovery of unique, robust, and specific enzyme catalysts. These enzymes are especially valuable for the cost-effective synthesis of chiral synthons for use in production of key pharmaceuticals, biological intermediates and natural product as well as for enzyme based combinatorial drug screening. The FACS screening system developed in Phase I handles 25,000 clones per second, incorporates assay methods for multiple enzyme types and realizes significant discovery rates. Building upon Phase I, Phase II efforts will be directed to refine and expand the screening capabilities for the FACS system to include several additional classes of enzymes based upon industrial needs for biocatalysts. In addition excision protocols will be improved to access environmental genomic libraries consisting of greater than 1 x 10 to the 8th power members and single cell PCR will be refined to capture DNA that encodes toxic products such as proteases.
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