Recombinant (2-->3)-alpha-sialyltransferase immobilized on nickel-agarose for preparative synthesis of sialyl Lewis(x) and Lewis(a) precursor oligosaccharides.

Tatiana Ivannikova (1) , Fabrice Bintein (1) , Annie Malleron (2) , Sylvie Juliant (2) , Martine Cerutti (2) , Anne Harduin-Lepers (3, 4) , Philippe Delannoy (3, 4) , Claudine Augé (1) , André Lubineau (1)

1 ICMMO - Institut de Chimie Moléculaire et des Matériaux d'Orsay
2 Baculovirus et Thérapie
3 Université Lille Nord de France (COMUE)
4 UGSF - Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576

The specificity of recombinant (2-->3)-alpha-sialyltransferase (ST3Gal-III), expressed in baculovirus-infected insect cells, has been determined with various oligosaccharide acceptors and sugar-nucleotide donors using a fluorescence based assay. Recombinant ST3Gal-III tagged with a polyhistidine tail was immobilized on Ni(2+)-NTA-Agarose as an active enzyme for use in the synthesis of three sialylated oligosaccharides: (i) the divalent molecule [alpha-Neu5Ac-(2-->3)-D-Galp-(1-->4)-beta-D-GlcpNAc-O-CH(2)](2)-C-(CH(2)OBn)(2) (12); (ii) the dansylated derivative, alpha-Neu5Ac-(2-->3)-D-Galp-(1-->3)-beta-D-GlcpNAc-O-(CH(2))(6)-NH-dansyl and; (iii) the tetrasacharide alpha-Neu5Ac-(2-->3)-beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-O-CH(3). Compound 12 was itself prepared from the divalent N-acetyllactosamine molecule built on pentaerythritol by a chemo-enzymatic route.

Domaines: Biochimie, Biologie Moléculaire

Tatiana Ivannikova, Fabrice Bintein, Annie Malleron, Sylvie Juliant, Martine Cerutti, et al.. Recombinant (2-->3)-alpha-sialyltransferase immobilized on nickel-agarose for preparative synthesis of sialyl Lewis(x) and Lewis(a) precursor oligosaccharides.. Carbohydrate Research, 2003, 338 (11), pp.1153-61. ⟨hal-01224169⟩