Heterologous expression of cis-prenyltransferase 1 for production of neryl pyrophosphate in Escherichia coli
Keywords:cis-prenyltransferase, neryl pyrophosphate, monoterpene synthase
Neryl pyrophosphate (NPP), also known as neryl diphosphate (NDP), is the cis isomer of genaryl pyrophosphate (GPP). Previously, it has been shown that monoterpene synthases found in plants use GPP as a substrate for the synthesis of monoterpenes and monoterpenoids. However, in 2009, Schilmiller identified a new monoterpene synthase gene for phellandrene synthase (PHS) that is expressed in glandular trichomes of the cultivated tomato Solanum lycopersicum and uses NPP as a substrate to produce β-phellandrene as a main product. An mRNA for a cis-prenyltransferase (cis-prenyltransferase 1, CPT1) was also identified in these glands. It encodes an enzyme that uses DMAPP and IPP to produce NPP. E. coli has the ability to synthesize GPP but not NPP. In this study, we cloned and expressed CPT1 in E. coli for production of NPP followed by coupling with a monoterpene synthase such as PHS, which specifically uses NPP as a substrate to produce β-phelandrene. Further metabolic engineering of the pathway and in situ product recovery would make this engineered E. coli a potential production platform for many valuable monoterpenes and their derivatives.