Glycosphingolipid synthesis employing a combination of recombinant glycosyltransferases and an endoglycoceramidase glycosynthase.

Glycosynthase mutants of Rhodococcus sp. endo-glycoceramidase II efficiently synthesize complex glycosphingolipids. Glycosyl fluoride donors may be assembled via sequential glycosyltransferase-catalysed glycosylation of lactosyl fluoride. Alternatively, lactosyl fluoride may be coupled to sphingosine prior to subsequent glycosylation steps.