Photosynthetic production of ethanol from carbon dioxide in genetically engineered cyanobacteria

The rapidly growing demand for energy and the environmental concerns about carbon dioxide emissions make the development of renewable biofuels more and more attractive. Tremendous academic and industrial efforts have been made to produce bioethanol, which is one major type of biofuel. The current production of bioethanol is limited for commercialization because of issues with food competition (from food-based biomass) or cost effectiveness (from lignocellulose-based biomass). In this report we applied a consolidated bioprocessing strategy to integrate photosynthetic biomass production and microbial conversion producing ethanol together into the photosynthetic bacterium, Synechocystis sp. PCC6803, which can directly convert carbon dioxide to ethanol in one single biological system. A Synechocystis sp. PCC6803 mutant strain with significantly higher ethanol-producing efficiency (5.50 g L−1, 212 mg L−1 day−1) compared to previous research was constructed by genetically introducing pyruvate decarboxylase from Zymomonas mobilis and overexpressing endogenous alcohol dehydrogenase through homologous recombination at two different sites of the chromosome, and disrupting the biosynthetic pathway of poly-β-hydroxybutyrate. In total, nine alcohol dehydrogenases from different cyanobacterial strains were cloned and expressed in E. coli to test ethanol-producing efficiency. The effects of different culturing conditions including tap water, metal ions, and anoxic aeration on ethanol production were evaluated.