L’objectif de ce projet de thèse était de caractériser le métabolisme de terpénoïdes (ou isoprénoïdes) chez les plantes supérieures. L’essentiel du travail a consisté à caractériser des triterpènes synthases (TTPS) d’Arabidopsis thaliana, un modèle végétal, ainsi que celles de l’épurge (Euphorbia lathyris), une euphorbe pour laquelle des applications agronomiques sont envisagées. Au cours de ma thèse, j’ai aussi contribué à l’étude du métabolisme et des fonctions des précurseurs de triterpènes et de stérols, ainsi qu’à leurs fonctions biologiques.Les triterpènes synthases, ou 2,3-oxydosqualène cyclases (OSCs), convertissent le substrat 2,3-oxydosqualene (SqO) en une multitude d'alcools triterpéniques, et ainsi amorcent la biosynthèse de dérivés triterpénoïdes (triterpènes oxydés, conjugués, etc ..). Arabidopsis thaliana contient 13 OSCs produisant divers squelettes triterpéniques, de type stéroïdien ou non-stéroïdien. Les produits de cyclisation du SqO ont été élucidés structuralement (GC-MS, RMN) après expression hétérologue des enzymes en levure erg7. Cette levure est déficiente en lanostérol synthase (ERG7), ce qui permet d'accumuler le SqO, substrat des cyclases. Lorsque le mutant est transformé avec un ADNc codant une triterpène synthase, il est capable de convertir le SqO en un ou plusieurs triterpènes. Cependant, la caractérisation des 13 OSCs d'Arabidopsis réalisée de façon hétérologue en levure n’a pas été établie inplanta. De façon surprenante, certains des composés produits dans les levures erg7 transformées n'ont jamais été détectés chez Arabidopsis. C'est pourquoi il a été nécessaire de reconsidérer les fonctions biochimiques exactes de ces enzymes dans un contexte végétal. / The subject of this PhD thesis is part of a research project entitled "Production of energy-rich triterpenoids in Euphorbia lathyris, a potential crop for third generation biofuels," whose acronym is EULAFUEL. This project is funded by a multipartner program ANR-KBBE and has been extended until December 2013. The aim of this PhD project is to get new insights into the aspects related with the biosynthesis and accumulation of latex triterpenoids. In addition, for comparison, a major objective of the thesis is to characterize functionally the enzymes involved in the synthesis of triterpenes in the model plant Arabidopsis thaliana. Triterpene synthases, also named oxidosqualene cyclases (OSCs), convert 2,3-oxidosqualene (OS) into a multitude of triterpene alcohols and there by initiate triterpene biosynthesis. Arabidopsis thaliana for instance has 13 OSCs producing diverse skeletons of steroidal or non-steroidal triterpenes. Cyclization products of a given enzyme have been characterized biochemically using a yeast heterologous expression system. However, for the majority of Arabidopsis triterpene synthases, inplanta studies are lacking. In fact, most of the compounds produced in yeast expressing such enzymes have never been detected in wild-type Arabidopsis. This is a reason why we should reconsider the exact biochemical function of triterpene synthases in the plant context. Then, in a comparative approach in E. lathyris, we project to study the specific triterpene accumulation in the laticifers, a specialized cell type where high amounts of lanosterol, an unusual OS cyclization product for plants, accumulate.

The transport sector is seen as particularly problematic when concerns about climate change and dependency on fossil energy are discussed. Because of this, bioenergy is strongly promoted for use in the transport sector, both on a European level and nationally in Sweden. Even though bioenergy is considered one of the key solutions, it is generally agreed that both supply- and demand-side measures will be needed to achieve a change to a more sustainable transport system. One of the reasons for this is the limited availability of biomass, especially agricultural feedstocks competing with food or feed production. Woody biomass, however more abundant, is also exposed to tough competition from other sectors. In this thesis, the role of biogas as a vehicle fuel in a future sustainable transport system is discussed together with the prerequisites needed to realise such a transport system. Biogas is a biofuel that could be produced in several different ways: by anaerobic digestion, which is a first-generation production route, by gasification, which is a second-generation process, and by catalytic reduction of carbon dioxide, a third-generation technology. The main focus in this thesis is on biogas produced by anaerobic digestion and the results show that there is a significant potential for an increase compared to today’s production. Biogas from anaerobic digestion, however, will only be able to cover a minor part of the demand in the Swedish transport sector. Considering biogas of the second and third generations, the potential for production is more uncertain in a mid-term future, mainly due to competition for feedstock, the possibility to produce other fuels by these processes, and the present immaturity of the technology. The limited potential for replacing fossil vehicle fuels, either by biogas or other renewable fuels, clearly shows the need for demand-side measures in the transport system as well. This thesis shows the importance of technical and non-technical means to decrease the demand for transport and to make the transport as efficient as possible. The results show that both energy-efficient vehicles and behavioural and infrastructural changes will be required. Policies and economic incentives set by governments and decision-making bodies have a prominent role to play, in order to bring about a shift to a more sustainable transport system, however, measures taken on individual level will also have a great impact to contribute to a more sustainable transport system. / <p>QC 20121116</p>