Domain SWapping of S-glycosyltransferases for the generation of multivalent THIO-glycoPEPtides – Sweet-Thio-Pep

The objective of the Sweet-THioPep project is to develop new biochemical tools as non-hydrolysable and multivalent probes to target lectins or glycosides hydrolases. These tools will be built by biocatalysis, using engineered glycosyltransferases (GT), modified by domain swapping, yielding chimerized GT, able to transfer a wide range of sugars onto thiols. This chimeric GT approach, based on bioinformatics analyses, is innovative, and will enable the generation of biocalaysts library, designed to graft diverse sugars on non-natural amino acids bearing thiol function. These amino acids will be incorporated in cyclic peptides (RAFT), to control their number and orientation, as well as providing the possibility to add othogonal functions. These scaffold will be then glycosylated by modified GT, yielding thiogycopeptides, coupling both multivalency and chemical and enzymatic stability of thioglycosides, never obtained so far using enzymatic biocatalysts. These multifunctionnal tools will eventually be used as lectin or enzyme probes in various applications ranging from cancer cells detection, to microbial pathogen anti-adhesive properties.