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Camptothecin (CPT) belongs to a group of monoterpenoidindole alkaloids (TIAs) and its derivatives such as irinothecan and topothecan have been widely used worldwide for the treatment of cancer, giving rise to rapidly increasing market demands. Genes from  Catharanthus roseus  encoding strictosidine synthase (STR) and geraniol 10-hydroxylase (G10H), were separately and simultaneously introduced into  Ophiorrhiza pumila  hairy roots. Overexpression of individual  G10H  (G lines) significantly improved CPT production with respect to non-transgenic hairy root cultures (NC line) and single  STR  overexpressing lines (S lines), indicating that  G10H  plays a more important role in stimulating CPT accumulation than  STR  in  O. pumila . Furthermore, co-overexpression of  G10H  and  STR  genes (SG Lines) caused a 56% increase on the yields of CPT compared to NC line and single gene transgenic lines, showed that simultaneous introduction of  G10H  and  STR  can produce a synergistic effect on CPT biosynthesis in  O. pumila . The MTT assay results indicated that CPT extracted from different lines showed similar anti-tumor activity, suggesting that transgenic  O. pumila  hairy root lines could be an alternative approach to obtain CPT. To our knowledge, this is the first report on the enhancement of CPT production in  O. pumila  employing a metabolic engineering strategy.

Co-overexpression of geraniol-10-hydroxylase and strictosidine synthase improves anti-cancer drug camptothecin accumulation in Ophiorrhiza pumila