Project

Project Overview

Alkanes, the primary component in diesel, are one class of molecules that cyanobacteria can synthesize. Furthermore, alkanes excreted from the cell then float on the surface of water, making harvesting of alkanes efficient and fairly simple through dewatering. Like other fatty-acid-derived products, acetyl-CoA is the initiation point for synthesis of alkanes. In a photosynthetic system such as cyanobacteria, acetyl-CoA is formed from pyruvate, which is the end product of glycolysis. Since pyruvate kinase catalyzes the final step in glycolysis (the conversion of phosphoenolpyruvate (PEP) to pyruvate), this enzyme is a common regulatory point of glycolysis in most organisms. Because the production of pyruvate provides the initial carbon source for lipid and alkane synthesis, we hypothesize that increasing the enzymatic activity of pyruvate kinase will increase the cellular concentration of pyruvate and thus increase the production of alkanes by the cell. In order to increase alkane production, we will insert a more activate pyruvate kinase gene in cyanobacteria (Synechocystis PCC 6803) in a broad host range plasmid (part BBa_K12500). The system could then be measured for pyruvate and alkanes through gas chromatography and a lipid profiling assay.