| 摘要 |
A reiterative method for obtaining the glycosyltransferases useful for glycosyltransferase-catalyzed synthesis of a saccharide composition by serially bonding preselected saccharide units onto an immobilized acceptor moiety which is one member selected from the group consisting of proteins, glycoproteins, lipids, glycolipids and carbohydrates. The method comprises in a first iteration, the steps of (i) contacting an acceptor moiety immobilized onto a solid or semi-solid support with a mixture containing at least one glycosyltransferase under conditions effective to allow said glycosyltransferase to bind to the immobilized acceptor moiety thereby resulting in an immobilized acceptor moiety-glycosyltransferase complex, followed by (ii) contacting the immobilized acceptor moiety-glycosyltransferase complex with a preselected saccharide under conditions and in the presence of co-reagents sufficient to effect bonding of the preselected saccharide unit onto the immobilized acceptor moiety catalyzed by the glycosyltransferase thereby freeing the glycosyltransferase from the complex and obtaining an immobilized product. This is followed by a single or a plurality of subsequent iterations of steps (i) and (ii), such that the immobilized product obtained in step (ii) of a given iteration is used as the immobilized acceptor moiety in step (i) of the following iteration. |
