Synthesis of 2,3-butanediol by Synechocystis sp. PCC6803 via heterologous expression of a catabolic pathway from lactic acid- and enterobacteria

Abstract

The direct and efficient conversion of CO2 into liquid energy carriers and/or bulk chemicals is crucial for a sustainable future of modern society. Here we describe the production of 2,3-butanediol in Synechocystis sp. PCC6803 expressing a heterologous catabolic pathway derived from enteric- and lactic acid bacteria. This pathway is composed of an acetolactate synthase, an acetolactate decarboxylase and an acetoin reductase. Levels of up to 0.72 g/l (corresponding to 8 mmol/L) of C(4) products, including a level of 0.43 g/l (corresponding to 4.7 mmol/L) 2,3-butanediol production are observed with the genes encoding these three enzymes integrated into the cyanobacterial genome, as well as when they are plasmid encoded. Further optimization studies revealed that Synechocystis expresses significant levels of acetolactate synthase endogenously, particularly under conditions of restricted CO2 supply to the cells. Co-expression of a soluble transhydrogenase or of an NADPH-dependent acetoin reductase allows one to drive the last step of the engineered pathway to near completion, resulting in pure meso-2,3-butanediol being produced.

Keywords: Butanediol; Catabolic pathway; Cell factory; Cyanobacteria; Metabolic engineering.