Patent ID: 7579173

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 and/or repetitive fashion to form extended glycosaminoglycan molecules, and wherein the at least one recombinant glycosaminoglycan transferase is selected from the group consisting of: (a) a recombinant glycosaminoglycan transferase having the amino acid sequence as set forth in SEQ ID NO: 4; (b) a recombinant glycosaminoglycan transferase encoded by a nucleotide sequence selected from the group consisting of SEQ ID NOs: 3 and 26; (c) a recombinant glycosaminoglycan transferase having an amino acid sequence encoded by a nucleotide sequence capable of hybridizing to the complement of at least one of SEQ ID NOS: 3 and 26 under hybridization conditions comprising hybridization at a temperature of 68° C. in 5x SSC/5x Denhardt's solution/1.0% SDS, followed with washing in 3x SSC at 42°; (d) a chimeric recombinant glycosaminoglycan transferase having an amino acid sequence selected from the group consisting of SEQ ID NOs: 74-76 and 85; (e) a recombinant glycosaminoglycan transferase having an amino acid sequence that is at least 90% identical to SEQ ID NO: 4; and (f) a recombinant glycosaminoglycan transferase encoded by a nucleotide sequence that is at least 90% identical to at least one of SEQ ID NOS: 3 and 26; and providing at least one UDP-sugar selected from the group consisting of UDP-GIcUA, UDP-GIcNAc, UDP-GIc, UDP-GaINAc, UDP-GIcN and UDP-GaIN 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 1.5, and wherein the desired size distribution is obtained by controlling the stoichiometric ratio of UDP-sugar to functional acceptor.