In Vivo Chaperone-Assisted Folding of α-1,6-Fucosyltransferase from Rhizobium sp.

Abstract

Glycosyltransferases have become powerful tools for the synthesis of oligosaccharides through their strict control over the stereo- and regioselectivity of glycosidic bond formation. One major drawback of glycosyltransferases in synthesis is their limited availability. The cloning and overexpression of bacterial glycosyltransferases is one alternative to overcome this problem. However, when a heterologous protein is over-expressed in E. coli misfolding and aggregation happen frequently, driving the recombinant protein into inactive aggregates known as inclusion bodies (I.B.). One possible and attractive strategy to avoid their formation is to increase the cellular levels of molecular chaperones. Chaperonins are able to mediate ATP-dependent folding of polypeptides to the native state. GroEL from E. coli is the best characterized chaperonin and its function is dependent on the cochaperonin GroES.