Patent ID: 6485930
Filing Date: 2002-11-26
Classification: C12N,C12P

Abstract:
A process for using mannosyl 1-phosphate (Man-1-P) to glycosylate an acceptor saccharide with mannose, with recycling of guanosine diphosphate that comprises the steps of:(a) admixing the following ingredients in an aqueous medium within a single vessel to form an aqueous reaction medium (i) mannosyl 1-phosphate; (ii) GDP-mannosyl pyrophosphorylase that catalyzes the formation of GDP-Man from the Man-1-P of (i) in the presence of GTP; (iii) a mannosyltransferase that forms a naturally occurring saccharide linkage; (iv) an acceptor saccharide for the mannosyl-transferase of (iii); and (v) a guanosine diphosphate recycling system that includes (a) GDP, GTP or both, (b) a phosphate donor, and (c) a kinase to transfer a phosphate group from the phosphate donor to GDP to form GTP, wherein each of the enzymes of (ii), (iii) and (v) is present in a catalytic amount; and (b) maintaining said aqueous reaction medium at a pH value of about 5 to about 10 at a temperature of about zero degrees C. to about 40Â° C. for a time period sufficient for said acceptor to be glycosylated.