Globally, Paclitaxel/Taxol® is an essential anticancer drug and there is a high demand for Taxol but there is a low yield from Taxus species. Two hundreds of reports have been shown for the production of taxol in endophytic fungi. Finding strategies for producing cost-effective microbial taxol is imperative to the demand. The endophytic fungi offer several advantages and the ability to produce Taxol easily with very less prices by fermentation technology, but a lack of information about the Taxol biosynthetic pathway, involved transcription factors and their industrial utilization has remained elusive. We have made objectives cloning of TB genes and TFs involved in fungal Taxol biosynthesis, In the this project, WRKY, AP2 ERF (ortholog to Taxus plant) bHLH and Zn(dt) (putative) TFs were obtained from two different endophytic fungal cDNA for the first time. Finding the elicitor-induced expression of fungal TFs and production of fungal taxol, our study finding noted that elicitors such as salicylic acid, methyl jasmonate (MeJA), sodium benzoate etc., have effectively stimulated taxol biosynthetic gene (HMGR, GGPPS, TDS, Tα5OH and PAL) expression and the enhanced production of fungal taxol, which was analyzed using qRT-PCR and various chromatography techniques, respectively. We have successfully overexpressed fungal TFs DgbHLH and PmERF into E. coli BL21 cells and purification of recombinant TFs from E. coli extracts, separately, the fungal TB gene promoters and determination of their DNA-binding sites to the TF proteins is underway. The study demonstrates that the TFs play significant roles in elicitor induced taxol biosynthesis from fungi. In addition, this project highlights that promising targets Transcription factors and Taxol biosynthetic genes for genetic and metabolic engineering approaches to further enhance fungal Taxol and precursors in future.
