Patent ID: 7741091

Claim:
A method for enzymatically producing a hyaluronic acid polymer in vitro, wherein the hyaluronic acid polymer is composed of less than about 150 sugars, comprising the steps of: providing a functional acceptor, wherein the functional acceptor has at least two sugar units selected from the group consisting of uronic acid and hexosamine; providing a hyaluronic acid synthase capable of elongating the functional acceptor, wherein the hyaluronic acid synthase is selected from the group consisting of: (a) a hyaluronic acid synthase having the amino acid sequence as set forth in SEQ ID NO:2; (b) a hyaluronic acid synthase encoded by the nucleotide sequence as set forth in SEQ ID NO:1; (c) a truncated form of (a) encoded by the nucleotide sequence as set forth in any of SEQ ID NOS:20, 27-30 and 71; (d) a truncated form of (a) having the amino acid sequence as set forth in SEQ ID NO:9; (e) a mutated form of (a) encoded by the nucleotide sequence as set forth in SEQ ID NO:42; and (f) a hyaluronic acid synthase encoded by a nucleotide sequence capable of hybridizing to the complement of the nucleotide sequence of SEQ ID NO:1 under hybridization conditions selected from the group consisting of: (i) 5×SSC/5× Denhardt's solution/1.0% SDS at 68° C., followed with washing in 0.2×SSC/0.1% SDS at room temperature; (ii) 1.8×HPB at about 30° C. to about 45° C. followed by washing in 0.2-0.5×HPB at about 45° C.; and (iii) 5×SSC/5× Denhardt's solution/1% SDS at 68° C., followed with washing in 3×SSC at 42° C.; and providing UDP-GlcUA and UDP-GlcNAc sugars such that the hyaluronic acid synthase elongates the functional acceptor to provide a hyaluronic acid polymer wherein the hyaluronic acid polymer is composed of less than about 150 sugars.