Testosteronan, a novel heparosan analog, testosteronan synthase, and methods of production and use thereof

A recombinant vector comprising an isolated nucleotide sequence encoding an enzymatically active testosteronan synthase wherein the testosteronan synthase is a single protein that is a dual-action catalyst that utilizes UDP-GlcUA and UDP-GlcNAc to synthesize a polysaccharide having the repeat structure [-4-D-GlcUA-α1,4-D-GlcNAc-α1-]n, the isolated nucleotide sequence comprising at least one of:

• (a) the nucleotide sequence of SEQ ID NO:2;
• (b) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1;
• (c) a nucleotide sequence that is at least 90% identical to SEQ ID NO:2;
• (d) a nucleotide sequence encoding an amino acid sequence that is at least 90% identical to SEQ ID NO:1;
• (e) a nucleotide sequence capable of hybridizing to a complement of SEQ ID NO:2 under hybridization conditions of at least one of:
• (1) 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
• (2) hybridization at a temperature of 30° C. in 5×SSC, 5×Denhardt's reagent, 30% formamide for about 20 hours followed by washing twice in 2×SSC, 0.1% SDS at about 30° C. for about 15 minutes followed by 0.5×SSC, 0.1% SDS at about 30° C. for about 30 minutes;
• (f) a nucleotide sequence capable of hybridizing to a complement of a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1 under hybridization conditions of at least one of (1) and (2) of (e) above;
• and
• (g) a nucleotide sequence encoding an amino acid sequence having up to 50 amino acid insertions, deletions or substitutions when compared to SEQ ID NO:1;
• and
• (h) a nucleotide sequence encoding an amino acid sequence having up to 25 amino acid insertions, deletions or substitutions when compared to SEQ ID NO:1.