Patent ID: 7223571

Claim:
A method for enzymatically producing defined glycosaminoglycan polymers comprising the steps of: providing at least one functional acceptor, wherein the functional acceptor has at least two sugar units selected from the group consisting of uronic acid and hexosamine; providing at least one recombinant glycosaminoglycan transferase having an empty acceptor site and being capable of elongating the at least one functional acceptor in a controlled fashion to form extended glycosaminoglycan molecules, the at least one recombinant glycosaminoglycan transferase selected from the group consisting of: (a) a recombinant glycosaminoglycan transferase having an amino acid sequence as set forth in SEQ ID NO:2; (b) a recombinant glycosaminoglycan transferase encoded by a nucleotide sequence as set forth in SEQ ID NO:1; (c) a truncated form of (a) encoded by a nucleotide sequence as set forth in any of SEQ ID NOS:10, 20, 27-32 and 71; (d) a mutated form of (a) encoded by a nucleotide sequence as set forth in any of SEQ ID NOS:11, 12, 16-19, 33-50; (e) a recombinant glycosaminoglycan transferase encoded by a nucleotide sequence capable of hybridizing to the nucleotide sequence of SEQ ID NO:1 under hybridization conditions comprising hybridization at a temperature of 68° C. in 5× SSC/5× Denhardt's solution/1.0% SDS, followed with washing in 3× SSC at 42° C.; and providing at least one UDP-sugar selected from the group consisting of UDP-GlcUA, UDP-GlcNAc, and UDP-GlcN in a stoichiometric ratio to the at least one functional acceptor such that the at least one recombinant glycosaminoglycan transferase elongates the at least one functional acceptor to provide glycosaminoglycan polymers wherein the glycosaminoglycan polymers have a desired size distribution such that the glycosaminoglycan polymers are substantially monodisperse in size such that the glycosaminoglycan polymers have a polydispersity value in a range of from 1.0 to 1.2, and wherein the desired size distribution is obtained by controlling the stoichiometric ratio of UDP-sugar to functional acceptor.