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2017-05-23 - Colloque/Présentation - communication orale - Anglais - 1 page(s)

Contreras Mogollon Angela , Leroy Baptiste , Mariage Pierre-Antoine, Wattiez Ruddy , "Optibiopharm: Powerful proteomic analysis of biosynthetic pathways involved in active compounds production in hairy roots of medicinal plants" in 3rd edition of international conferences : green chemistry and white biotech - industry of the future, Mons, Belgique, 2017

  • Codes CREF : Biochimie (DI3112), Phytopharmacie (DI3470), Biotechnologie (DI3800), Biologie moléculaire (DI3111), Biologie (DI3100)
  • Unités de recherche UMONS : Protéomie et Microbiologie (S828)
  • Instituts UMONS : Institut des Biosciences (Biosciences)

Abstract(s) :

(Anglais) Medicinal plants are known to possess a variety of bioactive compounds (secondary metabolites) that can be related to defence against different kind of stress and to a stop phase of the biomass growth. The large and diverse group of these metabolites includes alkaloids, anthraquinones, anthocyanins, flavonoids, saponins and terpenes, which can play an important role in cosmetics, nutritional and pharmaceuticals applications. Hairy roots culture of many of these plants have been established and shown to produce secondary metabolites more efficiently than natural roots in most cases, in addition, their use eliminates two major drawbacks of the use of plants on an industrial-scale, such as, food production and use of agricultural area, and to avoid dispersal of genetically modified organisms in the environment. Even if this method gives access to the production of active molecules with very high added value, an efficient and optimal use of this tool still requires significant increase in our understanding of this process. At present, most of the active molecules produced in hairy roots are induced, via an agent called elicitor. This elicitor may be a yeast extract or any molecule capable of inducing an equivalent stress. The modification of culture conditions is also often used. The elicitation triggers a stress which strongly disturbs the metabolism and stops roots’ growth. Optimisation of this process requires refinement in the elicitation strategy in order to trigger production of secondary metabolites while limiting undesirable effect on plants metabolism. In addition, the molecular study of biosynthetic pathways of these secondary metabolites can improve the knowledge of the production processes and also help in their optimisation. In this way, we are setting up a tray of analyses based on gel free differential proteomics, aiming at developing an experimental strategy that can be applied to any plant production model. We are focused on two particular cases, the ginsenosides production in Panax ginseng and the tanshinones production in Salvia miltiorrhiza, both compounds included in the group of terpenoids. Although many studies performed to date are related with the importance of these compounds, little is known about biochemical pathways involved in their production and regarding the regulation of those pathways. We have tested a variety of biotic and abiotic elicitors. HPLC analysis revealed an increase up to ten-fold of the production of the active compounds compared with non-elicited plants. We optimised proteins extraction strategy as well as LC MS/MS workflow and could identify more than 3200 different proteins (at least 2 unique peptides) in a single injection for S. milthiorriza and more than 2000 proteins for P. ginseng, using few mg of fresh material. Differential proteomic analysis ongoing will allow better understanding of the molecular mechanisms of biosynthesis of the active compounds in these two plants. Acknowledgements The research is supported by the Walloon region through the OptiBiofarm project (Beware Academia).