Taxol biosynthesis · De nova of transcription regulators from taxol®-producing endophytic fungi
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2021-09-01 → 2023-08-31
- EU contribution
- €174,806
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
De nova of transcription regulators from taxol®-producing endophytic fungi
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.
Data: CORDIS, © European Union
Project objective
SummaryWe propose to biosynthesis de novo of fungal taxol® production by the regulation of transcription factors (TFs). However, such a scenario is not yet to be investigated in case of taxol-producing endophytic fungi. The principle aim of this project is to isolate TFs genes WRKY, AP2, MYP etc. that specifically regulate the expression of taxol biosynthetic rate limiting gene promoters using transient RNAi. The attractive features of this study, examining the elicitor-induced regulator of selected TFs genes to see whether their mRNA levels correlate with fungal taxol biosynthesis. In order to identify key regulators of taxol pathway, cDNAs will be explore to obtain the recombinant TF protein. This project offer de nova interactions between the TFs and the TB gene promoters, which results of DNA binding cis-elements. It can be exploited to produce superior increased taxol yields from wild or transgenic fungi. This programme of work will provide a new strategy for unlocking the full potential of fungal biotechnology at the interface with metabolic engineering and rDNA technology to establish of fungal fermentation through genetic engineering approaches. The complementary expertise of Dr Kamal (Fungal biotechnology) and the supervisor Prof. Dr. Frank Kempken (FK) (Fungal genetic and molecular biology) offer the unique combination to realize the fungal taxol biosynthetic potential. Overall, this research will lead to a comprehensive understanding of the key TFs that control elicitor induced metabolic changes to produce highest yield of fungal taxol and will provide intellectual property (IP), technology development, which lead to Dr. Kamal with new and cutting-edge research training.
Original text from CORDIS.
Participants
- CHRISTIAN-ALBRECHTS-UNIVERSITAET ZU KIEL · KielCoordinatorGermany
Links
Data: CORDIS, © European Union