Browsing by Subject "Taxanes"

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    A rational approach to improving the biotechnological production of taxanes in plant cell cultures of Taxus spp.
    (Elsevier, 2014-03-27) Cusido, Rosa M.; Onrubia, Miriam; Sabater Jara, Ana Belén; Moyano, Elisabeth; Bonfill, Mercedes; Goossens, Alain; Pedreño, María Angeles; Palazón, Javier; Biología Vegetal
    Taxol is a complex diterpene alkaloid scarcely produced in nature and with a high anticancer activity. Biotechnological systems for taxol production based on cell cultures of Taxus spp. have been developed, but the growing commercial demand for taxol and its precursors requires the optimization of these procedures. In order to increase the biotechnological production of taxol and related taxanes in Taxus spp. cell cultures, it is necessary not only to take an empirical approach that strives to optimize in-put factors (cell line selection, culture conditions, elicitation, up-scaling, etc.) and out-put factors (growth, production, yields, etc.), but also to carry out molecular biological studies. The latter can provide valuable insight into how the enhancement of taxane biosynthesis and accumulation affects metabolic profiles and gene expression in Taxus spp. cell cultures. Several rational approaches have focused on studying the transcriptomic profiles of key genes in the taxol biosynthetic pathway in Taxus spp. cell cultures treated with elicitors such as methyl jasmonate, coronatine and cyclodextrins in relation with the taxane pattern, production and excretion to the culture medium. These studies have provided new insights into the taxol biosynthetic pathway and its regulation. Additionally, identifying genes with low levels of expression even in the presence of elicitors, together with metabolomics studies, has shed light on the limiting steps in taxol biosynthesis and could help define suitable metabolic targets for engineering with the main aim of obtaining highly productive Taxus cultured cells. In this review, we have summarized the latest endeavors to enhance the molecular understanding of the action mechanism of elicitors in Taxus spp. cell cultures. Developments in the ongoing search for new and more effective elicitation treatments and the application of metabolic engineering to design new transgenic cell lines of Taxus with an improved capacity for taxane production are described.
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    Transfecting Taxus x media protoplasts to study transcription factors BIS2 and TSAR2 as activators of taxane-related genes
    (Oxford University Press, 2020-03) Sánchez Muñoz, Raul; Almagro Romero, Lorena; Cusido, Rosa M.; Bonfill, Mercedes; Palazón, Javier; Moyano, Elisabeth; Biología Vegetal
    Taxane diterpenes are secondary metabolites with an im-portant pharmacological role in the treatment of cancer. Taxus spp. biofactories have been used for taxane produc-tion, but the lack of knowledge about the taxane biosynthet-ic pathway and its molecular regulation hinders their optimal function. The difficulties in introducing foreign genes in Taxus spp. genomes hinder the study of the mo-lecular mechanisms involved in taxane production, and a new approach is required to overcome them. In this study, a reliable, simple and fast method to obtain Taxus 􀀁 media protoplasts was developed, allowing their manipulation in downstream assays for the study of physiological changes in Taxus spp. cells. Using this method, Taxus protoplasts were transiently transfected for the first time, corroborating their suitability for transfection assays and the study of specific physiological responses. The two assayed transcription fac-tors (BIS2 and TSAR2) had a positive effect on the expression of several taxane-related genes, suggesting their potential use for the improvement of taxane yields. Furthermore, the results indicate that the developed method is suitable for obtaining T. 􀀁 media protoplasts for transfection with the aim of unraveling regulatory mechanisms in tax-ane production.