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Analysis of metabolites in the terpenoid pathway of Catharanthus roseus cell suspensions

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An Erratum to this article was published on 27 May 2015

Abstract

In Catharanthus roseus cell cultures, the monoterpenoid pathway has been shown to be a limiting factor in terpenoid indole alkaloid (TIA) production. This could be due to competition at the level of isopentenyl diphosphate::dimethylallyl diphosphate (C5) which leads to the biosynthesis of different terpenoid groups. For future engineering of the terpenoid pathway, chemical characterization of C. roseus cell cultures is a necessity. Therefore, in this study nine C. roseus cell suspension lines were characterized by analyzing the levels of the major terpenoids derived from different biosynthetic pathways which may compete for the same precursors; TIA (monoterpenoid, C10), carotenoids (tetraterpenoid, C40), and sterols (triterpenoid, C30). Among the cell lines, CRPP (S) was the most promising TIA-producing cell line which provided more TIA [24 μmol g−1 dry weight (DW)] than carotenoids (15 μmol g−1 DW) and sterols (2 μmol g−1 DW). However, when considering the distribution of the isopentenyl-precursor (C5), the carotenoids which assemble from 8× C5 represent twofold more C5-units (122 μmol g−1 DW) than the TIA in this cell line. In the CRPP (G), A12A2 (G), and A12A2 (S) cell lines, the C5 distribution was predominant toward carotenoid biosynthesis as well, resulting in a relatively high accumulation of carotenoids. The geranylgeranyl diphosphate (C20) pathway toward carotenoid production is therefore considered competitive toward TIA biosynthesis. For channeling more precursors to the TIA, the branch point for C10 and C20 seems an interesting target for metabolic engineering. Using principal component analysis of the chromatographic data, we characterized the cell lines chemically based on their metabolite levels. The information on the metabolic composition of C. roseus cell cultures is useful for developing strategies to engineer the metabolic pathways and for selection of cell lines for future studies.

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Acknowledgments

The authors thank Mr. L. Verhagen (Prisna B.V., Leiden, The Netherlands) for assistance with sterol analysis. The authors also thank the Ministry of Education, Malaysia and University of Malaya, Kuala Lumpur, Malaysia for financial support of Mohd Zuwairi Saiman. This research was funded by the IBOS-ACTS program as coordinated by NWO.

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Authors and Affiliations

  1. Natural Products Laboratory, Institute of Biology, Leiden University, 2300 RA, Leiden, The Netherlands

    Mohd Zuwairi Saiman, Natali Rianika Mustafa, Marianne Verberne, Robert Verpoorte, Young Hae Choi & Anna Elisabeth Schulte

  2. Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia

    Mohd Zuwairi Saiman

  3. ExPlant Technologies B.V., Galileiweg 8, 2333 BD, Leiden, The Netherlands

    Natali Rianika Mustafa, Barbora Pomahočová & Anna Elisabeth Schulte

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  1. Mohd Zuwairi Saiman
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  2. Natali Rianika Mustafa
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Correspondence to Natali Rianika Mustafa.

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Saiman, M.Z., Mustafa, N.R., Pomahočová, B. et al. Analysis of metabolites in the terpenoid pathway of Catharanthus roseus cell suspensions. Plant Cell Tiss Organ Cult 117, 225–239 (2014). https://doi.org/10.1007/s11240-014-0435-2

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