Quantification of metabolites in the indole alkaloid pathways ofCatharanthus roseus: Implications for metabolic engineering

Abstract

In this article, we present a review of the current state of metabolic engineering in Catharanthus roseus. A significant amount of research has contributed to characterization of several individual steps in the biosynthetic pathway of medicinally valuable alkaloids. However, knowledge of the regulation of these pathways is still sparse. Using hairy root cultures, we studied the responses of alkaloid metabolism to environmental stimulation such as light and elicitation. Through precursor feeding studies, the putative rate‐limiting steps of the terpenoid pathway in hairy root cultures also have been examined. Relating this knowledge to specific events at the molecular level, and the cloning of corresponding genes are the next key steps in metabolic engineering of the C. roseus alkaloids. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:333–338, 1998.

Non-systemic fungal endophytes in Carex brevicollis may influence the toxicity of the sedge to livestock

“…At first sight, these results suggest that the alkaloid is a plant product that can be induced by fungal endophytism. The synthesis of indole alkaloids, such as β-carbolines, elicited by fungal cell walls has been shown in plant cell cultures (Shanks et al, 1998;Facchini, 2001;Zhao et al, 2001;Bais et al, 2003;Pauw et al, 2004). Although a plant synthesis with a fungal regulation is plausible for brevicolline, more research is still needed since other explanations are possible.…”

Enhanced accumulation of catharanthine and vindoline in Catharanthus roseus hairy roots by overexpression of transcriptional factor ORCA2

“…However, vindoline, tabersonine as well as the dimeric alkaloids are restricted to leaves and stems because of the late steps of vindoline biosynthesis which require specialized cell types, idioblast and laticifer cells, which are located in stems and leaves (Westekemper et al, 1980;Deus-Neumann et al, 1987). Therefore, many cell and hairy root cultures produce catharanthine and tabersonine, but do not produce vindoline because there is a limitation in the conversion from tabersonine to vindoline (Shanks et al, 1998). This fact had been confirmed by some experiments (Bhadra and Shanks, 1997;Brillanceau et al, 1989;Toivonen et al, 1989).…”

Opium poppy and Madagascar periwinkle: model non‐model systems to investigate alkaloid biosynthesis in plants

“…Procedures have been developed for the transformation of opium poppy plants (Chitty et al, 2003;Facchini et al, 2008;Park and Facchini, 2000a), root cultures (Le Flem-Bonhomme et al, 2004;Park and Facchini, 2000b) and cell cultures (Belny et al, 1997). Stable transformation protocols for C. roseus are presently restricted to roots and cell cultures (Pasquali et al, 2006;Shanks et al, 1998;Zá rate and Verpoorte, 2007;Zhao and Verpoorte, 2007). Genetic transformation of opium poppy and C. roseus has provided the opportunity to alter the activity of individual enzymes of BIA biosynthesis, and to examine the consequences of such modifications on the accumulation of pathway products and intermediates.…”

“…Although tryptamine accumulation increased, the transgenic cell cultures remained limited in their ability to produce monoterpenoid precursors, such as loganin (van der Fits and Memelink, 2000). The potential of hairy root cultures for the stable production of alkaloids and other secondary metabolites has been attributed to a higher degree of genetic and biochemical stability due to increased cellular differentiation compared with cell cultures (Pasquali et al, 2006;Shanks et al, 1998;Zá rate and Verpoorte, 2007;Zhao and Verpoorte, 2007).…”