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On the way of synthetic biology of the furanocoumarin pathway
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Edité par CCSD -
International audience. Furanocoumarins are compounds commonly highlighted in 4 higher plant families: Rutaceae, Fabaceae, Moraceae, and Apiaceae. These molecules are allelochemicals involved in plant defense mechanisms against insects and phytopathogens. As many secondary metabolites, furanocoumarins are also of particular interest for therapeutic applications. Due to their physicochemical properties some of them are used for the treatment of skin diseases (bergapten) or cancers. The production of such molecule in high amounts might therefore be of particular interest.These compounds are synthesized through a complex cascade of enzymes involving cytochrome P450s, prenyltransferases, 2-oxoglutarate dependent dioxygenases and methyltransferases. Since 2007, most genes and enzymes involved in their biosynthesis have been described by our laboratory especially for the Apiaceae family. For this purpose we used several molecular biology strategies (published and non-published results).To develop an efficient furanocoumarin production system, we will overexpress the genes involved in the synthesis of these molecules in heterologous plant systems or in microorganisms. To achieve this goal all the genes identified so far will be cloned into a single plasmid using the newly described Goldenbraid technology, based on type IIS restriction enzymes.This challenging project will need to take into account the diversity of the enzymes involved in the biosynthetic pathway. Some enzymes were described to be cytosolic whereas others are known to be membrane bound. Some enzymes were highlighted in the cytoplasm whereas others are targeted to the endoplasmic reticulum or to chloroplasts. The expression level and the subcellular localization of each enzyme will also need to be optimized.
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