The limited bioavailability of plant-derived natural products with anticancer activity poses major challenges to the pharmaceutical industry.An example of this is camptothecin,a monoterpene indole alkaloid with potent...The limited bioavailability of plant-derived natural products with anticancer activity poses major challenges to the pharmaceutical industry.An example of this is camptothecin,a monoterpene indole alkaloid with potent anticancer activity that is extracted at very low concentrations from woody plants.Recently,camptothecin biosynthesis has been shown to become biotechnologically amenable in hairy-root systems of the natural producer Ophiorrhiza pumila.Here,time-course expression and metabolite analyses were performed to identify novel transcriptional regulators of camptothecin biosynthesis in O.pumila.It is shown here that camptothecin production increased over cultivation time and that the expression pattern of the WRKY transcription factor encoding gene OpWRKY2 is closely correlated with camptothecin accumulation.Overexpression of OpWRKY2 led to a more than three-fold increase in camptothecin levels.Accordingly,silencing of OpWRKY2 correlated with decreased camptothecin levels in the plant.Further detailed molecular characterization by electrophoretic mobility shift,yeast one-hybrid and dual-luciferase assays showed that OpWRKY2 directly binds and activates the central camptothecin pathway gene OpTDC.Taken together,the results of this study demonstrate that OpWRKY2 acts as a direct positive regulator of camptothecin biosynthesis.As such,a feasible strategy for the over-accumulation of camptothecin in a biotechnologically amenable system is presented.展开更多
INTRODUCTION Pentatrichomonas hominis,an anaerobic flagellated protozoan that inhabits the large intestines of mammals and belongs to the Trichomonadidae family(Wenrich 1944;Kim et al.2010;Li et al.2014b,2016,2018a,20...INTRODUCTION Pentatrichomonas hominis,an anaerobic flagellated protozoan that inhabits the large intestines of mammals and belongs to the Trichomonadidae family(Wenrich 1944;Kim et al.2010;Li et al.2014b,2016,2018a,2020;Maritz et al.2014;Zhang et al.2019),is mainly transmitted through the fecal-oral route.It was originally presumed to be a commensal protozoan(Tolbert et al.2012)but was found to cause gastrointestinal symptoms.展开更多
基金the National Key Research and Development Program of China(2018YFC1706203)National Natural Science Foundation of China(31571735,82073963,81522049,82003889)+3 种基金Zhejiang Provincial Ten Thousands Program for Leading Talents of Science and Technology Innovation(2018R52050)Zhejiang Provincial Program for the Cultivation of High-level Innovative Health Talents,Shanghai Science and Technology Committee Project(17JC1404300)Zhejiang Provincial Natural Science Foundation of China(LY20H280008,LQ21H280004)Zhejiang Chinese Medical University Research Foundation(2020ZR15).
文摘The limited bioavailability of plant-derived natural products with anticancer activity poses major challenges to the pharmaceutical industry.An example of this is camptothecin,a monoterpene indole alkaloid with potent anticancer activity that is extracted at very low concentrations from woody plants.Recently,camptothecin biosynthesis has been shown to become biotechnologically amenable in hairy-root systems of the natural producer Ophiorrhiza pumila.Here,time-course expression and metabolite analyses were performed to identify novel transcriptional regulators of camptothecin biosynthesis in O.pumila.It is shown here that camptothecin production increased over cultivation time and that the expression pattern of the WRKY transcription factor encoding gene OpWRKY2 is closely correlated with camptothecin accumulation.Overexpression of OpWRKY2 led to a more than three-fold increase in camptothecin levels.Accordingly,silencing of OpWRKY2 correlated with decreased camptothecin levels in the plant.Further detailed molecular characterization by electrophoretic mobility shift,yeast one-hybrid and dual-luciferase assays showed that OpWRKY2 directly binds and activates the central camptothecin pathway gene OpTDC.Taken together,the results of this study demonstrate that OpWRKY2 acts as a direct positive regulator of camptothecin biosynthesis.As such,a feasible strategy for the over-accumulation of camptothecin in a biotechnologically amenable system is presented.
基金This work was supported by the grants from the National Natural Science Foundation of China(No.32102696)the National Key Research and Development Program of China(No.2021YFF0702900).
文摘INTRODUCTION Pentatrichomonas hominis,an anaerobic flagellated protozoan that inhabits the large intestines of mammals and belongs to the Trichomonadidae family(Wenrich 1944;Kim et al.2010;Li et al.2014b,2016,2018a,2020;Maritz et al.2014;Zhang et al.2019),is mainly transmitted through the fecal-oral route.It was originally presumed to be a commensal protozoan(Tolbert et al.2012)but was found to cause gastrointestinal symptoms.
