Abstract: Geranyl pyrophosphate (GPP) is a key intermediate molecule in the bioproduction of thousands of natural products. Currently, natural products are either cultivated from plants, synthesized via complex chemical synthesis strategies, or through cell-based factories also known as biofoundries. However, in order to replicate the process in a cell free environment, numerous enzymes and cofactors must be utilized making this approach costly and unviable. In order to make this process viable, a new approach was needed that uses fewer enzymes and co-factors. As described herein, the present invention demonstrates that it is possible to create GPP from glycerol through a short and concise biosynthetic pathway outside of the cell.

Abstract: Glycerol is a byproduct of biodiesel and bioethanol production and its conversion to value-added chemicals is a promising avenue for realization of the biorefinery concept Conversion of glycerol to pyruvate through glycerate yields pyruvate, is a common intermediate of many high-value natural products. The present invention aims at improving the specific activity of a naturally occurring enzyme toward conversion of glycerate to pyruvate (TvDHT). The present invention features compositions of isolated dehydratase enzyme (DHT) polypeptide composition, in particular the DHT polypeptide composition comprising a single point mutation that increases the specific activity of the enzyme as compared to wild type.