One-pot multi-enzyme synthesis
As a large majority of enzymes function optimally near neutral pH, conditions for multi-enzyme one-pot oligosaccharide synthesis can often be identified. Glycosyltransferase based one-pot systems with the regeneration of sugar nucleotides have been reported for the synthesis of 6′-SLN (Ichikawa et al., 1991b), the α-Gal epitope (Figure 6A; Hokke et al., 1996; Fang et al., 1998), and a hyaluronic acid polymer (Figure 6B; De Luca et al., 1995). A one-pot synthesis of Lex using β1,4-GalT and α1,3-FucT has also been achieved (Arlt and Hindsgaul, 1995).
Furthermore, glycosidases and glycosyltransferases have been applied together in one-pot syntheses. In these cases, the glycosyltransferase removes the product from the glycosidase-catalyzed reaction, which drives the glycosidase equilibrium in the synthetic direction. This strategy has been employed in the synthesis of a core 2 trisaccharide (Dudziak et al., 1998), the sialyl-TF antigen (Kren and Thiem, 1995), and 6′-SLN (Herrmann et al., 1993).
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