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Background  A multi-monocistronic synthetic vector was used to assemble multiple genes of a nucleotide diphosphate (NDP)-sugar biosynthetic pathway to construct robust genetic circuits for the production of valuable flavonoid glycosides in  Escherichia coli . Characterized functional genes involved in the biosynthesis of uridine diphosphate (UDP)-glucose and thymidine diphosphate (TDP)-rhamnose from various microbial sources along with glucose facilitator diffusion protein ( glf ) and glucokinase ( glk ) from  Zymomonas mobilis  were assembled and overexpressed in a single synthetic multi-monocistronic operon.    Results  The newly generated NDP-sugars biosynthesis circuits along with regiospecific glycosyltransferases from plants were introduced in  E. coli  BL21 (DE3) to probe the bioconversion of fisetin, a medicinally important polyphenol produced by various plants. As a result, approximately 1.178 g of fisetin 3- O -glucoside and 1.026 g of fisetin 3- O -rhamnoside were produced in UDP-glucose and TDP-rhamnose biosynthesis systems respectively, after 48 h of incubation in 3 L fermentor while supplementing 0.9 g of fisetin. These yields of fisetin glycosides represent ~99% of bioconversion of exogenously supplemented fisetin. The systems were also found to be highly effective in bio-transforming other flavonols (quercetin, kaempferol, myricetin) into their respective glycosides, achieving over 95% substrate conversion.    Conclusion  The construction of a synthetic expression vector for bacterial cell factory followed by subsequent re-direction of metabolic flux towards desirable products have always been revolutionized the biotechnological processes and technologies. This multi-monocistronic synthetic vector in a microbial platform is customizable to defined task and would certainly be useful for applications in producing and modifying such therapeutically valued plant secondary metabolites.    Electronic supplementary material  The online version of this article (doi:10.1186/s12934-015-0261-1) contains supplementary material, which is available to authorized users.

Synthetic sugar cassettes for the efficient production of flavonol glycosides in Escherichia coli