Biotransformations Near and Above 100 C: Mining Genomes of Hyperthermophiles for New Biocatalysts

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Biocatalyst discovery and characterization remains a key biotechnological objective. High throughput screening and directed evolution approaches have been developed over the last few years that enable such biocatalysts to be identified and then optimized with respect to their biochemical, biophysical and bioprocessing properties. However, current methods for discovery and optimization rely heavily on enzyme-based assays, which may be difficult or even infeasible to implement because of chemical limitations or gene expression issues. Moreover, multienzyme transformations are not easily identified by this approach since current methods focus on the products of individual genes. As a means of circumventing the limitations with current approaches, the objective of this research is to develop functional genomics-based discovery methods to identify novel biocatalysts. With the advent of high-throughput technologies to measure gene expression, cellular responses at the genetic level can be correlated to environmental conditions. This alternative approach to enzyme discovery takes advantage of highly sensitive, transcriptional.based cellular responses to biotechnologically-related environmental challenges and growth conditions. In this project period, extensive use will be made of DNA microarrays to identify the genes that encode new enzymes for a range of important biotransformations.