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Theses

New computational approaches for investigating the impact of mutations on the transglucosylation activity of sucrose phosphorylase enzyme

Abstract : In this thesis, we explore the usage of computational approaches for understanding the link between mutations and changes in protein activity. Our study model is a bacterial sucrose phosphorylase enzyme from Bifidobacterium adolescentis (BaSP). This glycosyl hydrolase from family 13 (GH13) has been a focus in the industry due to its ability to synthesize original disaccharides and glycoconjugates. In fact, its activity is to transfer a glucose moiety from a donor sucrose to an acceptor which can be a monosaccharide or a hydroxylated aglycone. The enzymatic reaction proceeds by a double displacement with retention of configuration mechanism whereby a covalent glucosyl-enzyme intermediate is formed. However, it is at stake to control the regioselectivity of this transfer for it to be applicable at industrial level. This thesis aimed at providing a rational explanation for the observed impact of mutations on the regioselectivity of BaSP in view of controlling the synthesis of rare pre-biotic disaccharides like kojibiose and nigerose. We hypothesized that the preferred orientations of the acceptor determines the regioselectivity of the enzyme. In that respect, we used computational approaches to investigate the impact of mutations on the binding of the acceptor to the glucosyl-enzyme intermediate. The methodology used in this work opens the perspective of using computational approaches for engineering the regioselectivity of of glycosyl hydrolases with similar mechanism.
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Mahesh Velusamy. New computational approaches for investigating the impact of mutations on the transglucosylation activity of sucrose phosphorylase enzyme. Bioinformatics [q-bio.QM]. Université de la Réunion, 2018. English. ⟨NNT : 2018LARE0045⟩. ⟨tel-02874581⟩

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