Sequential hydrolysis of hyaluronate by beta-glucuronidase and beta-N-acetylhexosaminidase.
1. Hyaluronate extracted from rooster comb was digested by a mixture of beta-N-acetylhexosaminidase and beta-glucuronidase with simultaneous dialysis for 96 h. 2. The produjct, yielding 99.6% of a mixture of mono- and oligo-saccharides, was purified by gel chromatography and analysed for glucuronic acid, N-acetylglucosamine and other sugars. 3. The oligosaccharide portion was chromatographed on DEAE-cellulose, and the effluent fractions were analysed for glucuronic acid and N-acetylglucosamine, reduced with NaBH4, digested by beta-N-acetylhexosaminidase and subjected to acid hydrolysis and glucosamine determination. 4. GlcNAc-GlcA-GlcNAc, GlcA-GlcA-GlcNAc and GlcA-GlcA-GlcA-GlcNAc were the oligosaccharides obtained, which resulted from the transferase activity of the enzymes and represented 57% of the digestion products. The results demonstrate that this hyaluronate is an unbranched polymer of approximatey equal amounts of glucuronic acid and N-acetylglucosamine. The data also indicate that if this glycosaminoglycan contains any of the neutral sugars for which it was analysed, their concentration must be less than 0.020% of the sum of the known components.
D000117:Acetylglucosamine / Q000032:analysis; D000818:Animals; D002645:Chickens; D002850:Chromatography, Gel; D002855:Chromatography, Thin Layer; D003129:Comb and Wattles / Q000032:analysis; D005965:Glucuronates / Q000032:analysis; D020723:Glucuronic Acid; D005966:Glucuronidase; D006596:Hexosaminidases; D006820:Hyaluronic Acid; D006868:Hydrolysis; D008297:Male; D009844:Oligosaccharides / Q000032:analysis; D001619:beta-N-Acetylhexosaminidases
D005965:Glucuronates; D009844:Oligosaccharides; D020723:Glucuronic Acid; D006820:Hyaluronic Acid; D006596:Hexosaminidases; D005966:Glucuronidase; D001619:beta-N-Acetylhexosaminidases; D000117:Acetylglucosamine