Suspension cultured transgenic cells of Nicotiana tabacum expressing tryptophan decarboxylase and strictosidine synthase cDNAs from Catharanthus roseus produce strictosidine upon secologanin feeding

Hallard, Didier; Heijden, Robert van der; Verpoorte, Robert; Cardoso, Inês Lopes; Pasquali, Giancarlo; Memelink, Johan; Hoge, J. H. C.

doi:10.1007/s002990050350

Cited by 46 publications

(20 citation statements)

References 16 publications

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“…While studies involving genetic engineering of plant secondary metabolite profiles are becoming more common, very few transgenic plant cell cultures have been used in biotransformations. In one case, transgenic tobacco cells expressing foreign tryptophan decarboxylase (TDC) and strictosidine synthase (STR) enzymes produced strictosidine (a glucosylated alkaloid intermediate) when secologanin (STR substrate) was fed to the cell cultures (Hallard et al 1997). In this study, while the STR activity was only found inside the cells, the majority of the strictosidine accumulated in the culture medium.…”

Section: Resultsmentioning

confidence: 97%

Biotransformation of an exogenously supplied isoflavonoid by transgenic tobacco cells expressing alfalfa isoflavone reductase

2002

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Isoflavone reductase (IFR) from alfalfa catalyzes the NADPH-dependent reduction of 2′-hydroxy isoflavones to 2′-hydroxyisoflavanones such as vestitone, thereby performing a key step in isoflavonoid phytoalexin biosynthesis. Tobacco (Nicotiana tabacum L. cv. Xanthi) was transformed with an alfalfa cDNA encoding IFR regulated by an enhanced cauliflower mosaic virus 35S promoter and used to generate cell suspension cultures. Transformed cells expressed high levels of IFR activity and could take up 2′-hydroxyformononetin (2′OHF; IFR substrate) and convert it to vestitone. Both 2′OHF and vestitone were each converted to two sugar conjugates. The structure of the most abundant vestitone conjugate was determined using NMR spectroscopic analysis to be a novel vestitone 2′-O-glucoside. Liquid chromatography-mass spectroscopy (LC-MS) indicated that the molecular weights of the three less abundant conjugates were consistent with hexose conjugates of vestitone and 2′OHF. The IFR reduction and conjugation reactions were complete within 4 h, with the net percentage conversion of 2′OHF to vestitone ranging as high as 60%.

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Section: Resultsmentioning

confidence: 97%

Biotransformation of an exogenously supplied isoflavonoid by transgenic tobacco cells expressing alfalfa isoflavone reductase

2002

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“…Since these ABC transporters and H + -antiporters are less specific in substrate preference and highly regulated (Yazaki 2005); their activity definitely affects production of indole alkaloids in C. roseus cell cultures. Overexpression of indole alkaloidbiosynthesis genes in tobacco cells showed that strictosidine generated by transgenic tobacco cell cultures fed with secologanin and tryptamine are exported into the medium and not stored in the vacuole (Hallard et al 1997;Verpoorte, unpublished results). Apparently every plant species has different selective transport systems.…”

Section: Biosynthetic and Metabolic Pathway Gene And Enzyme Charactementioning

confidence: 95%

“…The regulation includes probably substrate and product trafficking, compartmentation and metabolism, since transgenic cells still kept high enzyme activity. The results from transgenic tobacco and Cinchona officinalis hairy roots expressing the TDC and STR genes also point to the importance of subcellular trafficking and storage for production of secondary metabolites (Hallard et al 1997;.…”

Section: Manipulating Biosynthetic Genesmentioning

confidence: 97%

Manipulating indole alkaloid production by Catharanthus roseus cell cultures in bioreactors: from biochemical processing to metabolic engineering

2007

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Catharanthus roseus plants produce many pharmaceutically important indole alkaloids, of which the bisindole alkaloids vinblastine and vincristine are antineoplastic medicines and the monoindole alkaloids ajmalicine and serpentine are antihypertension drugs. C. roseus cell cultures have been studied for producing these medicines or precursors catharanthine and vindoline for almost four decades but so far without a commercially successful process due to biological and technological limitations. The research thus focused on the one hand on engineering the bioreactor process on the other engineering the cell factory itself. 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 alkaloid production in recent years. The paper also attempts to highlight new strategies and technologies to improve alkaloid production and bioreactor performance. Perspectives of metabolic engineering to create new cell lines for large-scale production of indole alkaloids in bioreactors and effective combination of these up-and downstream processing are presented.

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“…In particular the Tdc and Str genes have been extensively studied in C. roseus cultures as mentioned above (Canel et al 1998;Whitmer et al 2002). These genes have also been expressed heterologously in, for instance, tobacco cells (Hallard et al 1997) where tryptamine was produced from tryptophan by TDC enzyme, and upon feeding with secologanin, strictosidine was also formed, catalysed by the STR enzyme. However, strictosidine was not further metabolised since tobacco cells lacked glucosidase activity.…”

Section: Manipulation Of Developmentally Regulated Genes Homologous mentioning

confidence: 98%

Strategies for the genetic modification of the medicinal plant Catharanthus roseus (L.) G. Don

Zárate

Verpoorte

2007

Phytochem Rev

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This review describes the different plant transformation techniques, including guided infection with Agrobacterium tumefaciens and A. rhizogenes, particle bombardment and protoplast fusion, that have been attempted to create transgenic Catharanthus roseus (L.) G. Don cell cultures, hairy roots and whole plants. The review also focuses on the different approaches used to manipulate and improve secondary metabolite yields in various culture systems, with special attention to the most relevant results achieved. Finally, under future perspectives, the authors propose several approaches which would likely be implemented with this species, to try to boost the accumulation of the anti-tumour agents, vinblastine and vincristine. Some comments on how the future of the genetic manipulation of medicinal plants may proceed aiming at achieving higher secondary metabolite yields are also given.

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Suspension cultured transgenic cells of Nicotiana tabacum expressing tryptophan decarboxylase and strictosidine synthase cDNAs from Catharanthus roseus produce strictosidine upon secologanin feeding

Cited by 46 publications

References 16 publications

Biotransformation of an exogenously supplied isoflavonoid by transgenic tobacco cells expressing alfalfa isoflavone reductase

Biotransformation of an exogenously supplied isoflavonoid by transgenic tobacco cells expressing alfalfa isoflavone reductase

Manipulating indole alkaloid production by Catharanthus roseus cell cultures in bioreactors: from biochemical processing to metabolic engineering

Strategies for the genetic modification of the medicinal plant Catharanthus roseus (L.) G. Don

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