Heterologous synthesis of geranyllinalool, a diterpenol plant product, in the cyanobacterium Synechocystis

Abstract

Cyanobacteria are industrially robust photosynthetic microorganisms that can be genetically programmed to synthesize commodity products for domestic and industrial consumption. In the present work, Synechocystis was endowed with the synthesis of the plant secondary metabolite geranyllinalool, a diterpene alcohol of commercial interest. Total average yields of 360 μg of geranyllinalool per gram of dry cell weight were obtained in the course of a 48-h cultivation period. Geranyllinalool was primarily sequestered inside the transformant cells, corresponding to 60-70% of the total heterologous product, instead of being entirely exuded, as the case is with shorter heterologous terpene hydrocarbons. Extraction of geranyllinalool necessitated disruption of the cells in order to release and isolate this chemical product. Moreover, geranyllinalool accumulation in the cells caused a mild inhibitory effect on cell fitness and biomass growth rate, such that the duplication time of Synechocystis transformants was 1.4-fold longer than that of the control. The remaining 30-40% of the geranyllinalool product was found to float on the surface of sealed transformant cultures, where it was siphoned off by applying a hydrophobic overlayer, with no need to disrupt the cells in this case. Concluding, the work extended efforts to heterologously produce terpene and terpenol products in cyanobacteria, and addressed possibilities and constrains inherent to this production system.

Keywords: Cyanobacteria; Diterpene alcohol; Geranyllinalool; Metabolic engineering; Nicotiana attenuata; Terpenoids.