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Keywords:

  • biocatalysis;
  • carbohydrates;
  • cis-glycosylation;
  • cyclodextrin glucanotransferase;
  • Shigella flexneri

Abstract

We report the enzymatic synthesis of α-D-glucopyranosyl-(1[RIGHTWARDS ARROW]4)-α-L-rhamnopyranoside and α-D-glucopyranosyl-(1[RIGHTWARDS ARROW]3)-α-L-rhamnopyranoside by using a wild-type transglucosidase in combination with glucoamylase and glucose oxidase. It was shown that Bacillus circulans 251 cyclodextrin glucanotransferase (CGTase, EC 2.1.4.19) can efficiently couple an α-L-rhamnosyl acceptor to a maltodextrin molecule with an α-(1[RIGHTWARDS ARROW]4) linkage, albeit in mixture with the α-(1[RIGHTWARDS ARROW]3) regioisomer, thus giving two glucosylated acceptors in a single reaction. Optimisation of the CGTase coupling reaction with β-cyclodextrin as the donor substrate and methyl or allyl α-L-rhamnopyranoside as acceptors resulted in good conversion yields (42–70 %) with adjustable glycosylation regioselectivity. Moreover, the efficient chemical conversion of the products of CGTase-mediated cis-glucosylation into protected building blocks (previously used in the synthesis of O-antigen fragments of several Shigella flexneri serotypes) was substantiated. These novel chemoenzymatic strategies towards useful, convenient intermediates in the synthesis of S. flexneri serotypes 2a and 3a oligosaccharides might find applications in developments towards synthetic carbohydrate-based vaccine candidates against bacillary dysentery.