Journal ArticleDOI
Effect of precursor feeding on alkaloid accumulation by a strictosidine synthase over-expressing transgenic cell line S1 of Catharanthus roseus
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The results suggest that high rates of tryptamines synthesis can still take place under conditions of low TDC activity and the flux towards tryptamine is induced by loganin feeding, however, accumulation oftryptamine seems to reduce the flux through feedback inhibition.Abstract:
The transgenic S1 cell line of Catharanthus roseus (L.) G. Don has been used to study possible rate limiting steps in the terpenoid indole alkaloid (TIA) biosynthesis. Line S1 carries a recombinant, over-expressed version of the endogenous Str gene which encodes strictosidine synthase (STR; EC 4.3.3.2). STR catalyzes the stereospecific condensation of tryptamine and secologanin to strictosidine. Various concentrations and combinations of biosynthetic indole precursors L-tryptophan, tryptamine, and iridoid precursors loganin and secologanin were added to the cell suspension cultures of line S1. The largest TIA accumulation occurred when the precursor was supplied at the time of inoculation of the cells into the production medium. Line S1 could supply tryptamine endogenously up to 0.8 mM loganin feeding. The enhancement of the accumulation of TIAs by addition of loganin indicates a limitation in the terpenoid pathway. Supplying tryptamine or tryptophan along with the iridoid precursors resulted in even further increase of alkaloid accumulation. Under optimal conditions, cultures of line S1 accumulated about 600 μmol l−1 of TIAs. Also, the conversion of strictosidine into other TIAs further down the pathway seems to be a limiting step. Considering the mass balance of the intermediates fed and TIAs recovered, several yet unknown pathways must be involved in channeling away intermediates from the TIA pathway and in the breakdown of the TIAs. Our results suggest that high rates of tryptamine synthesis can still take place under conditions of low TDC activity and the flux towards tryptamine is induced by loganin feeding. However, accumulation of tryptamine seems to reduce the flux through feedback inhibition.read more
Citations
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Journal ArticleDOI
Gene-to-metabolite networks for terpenoid indole alkaloid biosynthesis in Catharanthus roseus cells
Heiko Rischer,Matej Orešič,Tuulikki Seppänen-Laakso,Mikko Katajamaa,Freya Lammertyn,Wilson Ardiles-Diaz,Marc Van Montagu,Dirk Inzé,Kirsi-Marja Oksman-Caldentey,Alain Goossens +9 more
TL;DR: A comprehensive profiling analysis of the Madagascar periwinkle (Catharanthus roseus), a source of the anticancer drugs vinblastine and vincristine, provides the basis for a better understanding of periwinkles secondary metabolism and increases the practical potential of metabolic engineering of this important medicinal plant.
Journal ArticleDOI
Catharanthus terpenoid indole alkaloids: biosynthesis and regulation
TL;DR: Most of the work focussed on early steps of the pathway, particularly the discovery of the 2-C-methyl-d-erythritol 4-phosphate (MEP)-pathway leading to terpenoids, and much progress has been made at alkaloid regulatory level.
Journal ArticleDOI
Indole alkaloids from Catharanthus roseus: bioproduction and their effect on human health.
TL;DR: This review is focused on the different strategies which improve TIA production, and in the analysis of the beneficial effects that these compounds exert on human health.
Journal ArticleDOI
Transcriptional response of the terpenoid indole alkaloid pathway to the overexpression of ORCA3 along with jasmonic acid elicitation of Catharanthus roseus hairy roots over time.
TL;DR: This work explores the transient effects of overexpressing ORCA3 under the control of a glucocorticoid-inducible promoter system in C. roseus hairy roots along with the simultaneous feeding of JA, and finds that JA treatment caused the largest increase in TIA metabolites and transcripts of pathway genes.
Journal ArticleDOI
Manipulating indole alkaloid production by Catharanthus roseus cell cultures in bioreactors: from biochemical processing to metabolic engineering
Jian Zhao,Robert Verpoorte +1 more
TL;DR: This review mainly summarizes the progress made on biochemical engineering aspects of C. roseus cell cultures in bioreactors in the past decades and metabolic engineering of indole alkaloids production in recent years and attempts to highlight new strategies and technologies to improve alkaloid production and bioreactor performance.
References
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Journal ArticleDOI
A revised medium for rapid growth and bio assays with tobacco tissue cultures
Toshio Murashige,Folke Skoog +1 more
TL;DR: In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provide a basis for understanding how compartment-specific redox dynamics may operate in retrograde signaling and stress 67 acclimation in plants.
Book ChapterDOI
Formation of the Indole Alkaloids Serpentine and Ajmalicine in Cell Suspension Cultures of Catharanthus roseus
TL;DR: Since there is an overall shortage of Rauwolfia drugs, increasing use is being made of the serpentine reduction method, and in roots of the fast growing Catharanthus roseus low amounts of ajmalicine, but relatively large amounts of serpentine are produced.
Journal ArticleDOI
Effects of over-expression of strictosidine synthase and tryptophan decarboxylase on alkaloid production by cell cultures of Catharanthus roseus
Camilo Canel,M. Inês Lopes-Cardoso,Serap Whitmer,Leslie van der Fits,Giancarlo Pasquali,Robert van der Heijden,J. Harry C. Hoge,Robert Verpoorte +7 more
TL;DR: It is concluded that constitutive over-expression of Str is highly desirable for increased TIA production, however, given its complexity, limited intervention in the TIA pathway will yield positive results only in the presence of a favorable epigenetic environment.
Journal ArticleDOI
Effect of terpenoid precursor feeding and elicitation on formation of indole alkaloids in cell suspension cultures of Catharanthus roseus
TL;DR: The effects of terpenoid precursor feeding and elicitation by a biotic elicitor on alkaloid production of Catharanthus roseus suspension cultures were studied and increased tryptamine accumulation in non- fed cells but it did not further increase alkaloids accumulation in precursor-fed cells.