title: Novel products in hyaluronan digested by bovine testicular hyaluronidase.
abstract: When the products of hyaluronan (HA) digested by bovine testicular hyaluronidase (BTH) were analyzed by high-performance liquid chromatography (HPLC), minor peaks were detected just before the main even-numbered oligosaccharide peaks. The amount of each minor peak was dependent on the reaction conditions for transglycosylation, rather than hydrolysis, by the BTH. Mainly based on HPLC and MS analysis, each minor peak was found to correspond to its oligosaccharide with one N-acetyl group removed from the reducing terminal N-acetylglucosamine. Enzymatic studies showed that the N-deacetylation activity was closely related to reaction temperature, pH, and the concentration of NaCl contained in the buffer, and glycosaminoglycan types and chain lengths of substrates. These findings strongly suggest that the N-deacetylation reaction in minor peaks was due to a novel enzyme contaminant in the BTH, N-deacetylase, that carries out N-deacetylation at the reducing terminal N-acetylglucosamine of oligosaccharides and is dependent on HA hydrolysis by BTH.
authors.affiliation: Department of Glycotechnology, Center for Advanced Medical Research, Hirosaki University Graduate School of Medicine, Hirosaki, Japan. seika1@cc.hirosaki-u.ac.jp
authors.forename: Fengchao
authors.initials: F
authors.lastname: Chen
authors.forename: Ikuko
authors.initials: I
authors.lastname: Kakizaki
authors.forename: Masanori
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Kaoru
authors.initials: K
authors.lastname: Kojima
authors.forename: Keiichi
authors.initials: K
authors.lastname: Takagaki
authors.forename: Masahiko
authors.initials: M
authors.lastname: Endo
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D006820:Hyaluronic Acid / Q000737:chemistry / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008297:Male; D013058:Mass Spectrometry; D015394:Molecular Structure; D013696:Temperature; D013737:Testis / Q000201:enzymology*
chemical_list: D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase