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Theses

Conception de nouveaux biocatalyseurs par fusion de domaines catalytiques

Abstract : Microbial production of molecules of interest can be improved by severa! engineering strategies. Enzymatic engineering has a central role in the conception of efficient host because pathway's efficiency depends in first place on the efficiency of the enzymes. Knowing which synthetic enzymes conceptions are efficient and knowing to characterize the best candidates are essential. Enzyme colocalisation inside metabolic pathway might improve the production of final molecule of interest by allowing rapid biotransformation of intermediates of the pathway. Multidomain proteins regrouping severa! enzymatic activities are described in the literature. This work has focused in part on the creation of synthetic fusion of sorne carotenogenic enzymes for the production of beta carotene in Saccharomyces cerevisiae. Different types of enzymatic fusions and configurations have been tested and. characterized. The study allowed the creation of an efficient tripartite enzyme. fusion which produces two times Jess intermediates and two times more beta carotene. Precise measurement of each caro teno id' s concentration coupled with quantification of enzymes allows the characterization of the efficiency of each synthetic enzyme. Other strategies for enzyme spatial co localisation have also been tested using domains of interaction like cohesin-dockerin or the oligomeric protein CcmK2. Sorne carotenogenic enzymes are still functional using those configurations. Sorne of the enzymatic systems modify the metabolic flow ofcarotenoids and produce carotenoids different from the natural systems. Sorne strategies have changed the metabolic flux of carotenoids inside the pathway. Interestingly, a fine control of activity of enzyme might allow a fine control of the nature of the final carotenoid
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Submitted on : Friday, January 17, 2020 - 9:15:09 AM
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  • HAL Id : tel-02443294, version 1

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Mamy Hery Ny Aina Rabeharindranto. Conception de nouveaux biocatalyseurs par fusion de domaines catalytiques. Microbiologie et Parasitologie. INSA de Toulouse, 2019. Français. ⟨NNT : 2019ISAT0015⟩. ⟨tel-02443294⟩

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