title: Improvement of the digestibility of sulfated hyaluronans by bovine testicular hyaluronidase: a UV spectroscopic and mass spectrometric study.
abstract: Glycosaminoglycans (GAGs) such as hyaluronan (HA) and chondroitin sulfate (CS) are important, natural polysaccharides which occur in biological (connective) tissues and have various biotechnological and medical applications. Additionally, there is increasing evidence that chemically (over)sulfated GAGs possess promising properties and are useful as implant coatings. Unfortunately, a detailed characterization of these GAGs is challenging: although mass spectrometry (MS) is one of the most powerful tools to elucidate the structures of (poly)saccharides, MS is not applicable to high mass polysaccharides, but characteristic oligosaccharides are needed. These oligosaccharides are normally generated by enzymatic digestion. However, chemically modified (particularly sulfated) GAGs are extremely refractive to enzymatic digestion. This study focuses on the investigation of the digestibility of GAGs with different degrees of sulfation by bovine testicular hyaluronidase (BTH). It will be shown by using an adapted spectrophotometric assay that all investigated GAGs can be basically digested if the reaction conditions are carefully adjusted. However, the oligosaccharide yield correlates reciprocally with the number of sulfate residues per polymer repeating unit. Finally, matrix-laser desorption and ionization (MALDI) MS will be used to study the released oligosaccharides and their sulfation patterns.
authors: Institute of Medical Physics and Biophysics, Medical Faculty, University of Leipzig, Härtelstraße 16-18, 04107 Leipzig, Germany.
authors: Katharina
authors: 0000-0001-9328-7319
authors: K
authors: Lemmnitzer
authors: Institute of Medical Physics and Biophysics, Medical Faculty, University of Leipzig, Härtelstraße 16-18, 04107 Leipzig, Germany.
authors: Jürgen
authors: 0000-0002-4328-8652
authors: J
authors: Schiller
authors: INNOVENT e. V., Biomaterials Department, Pruessingstraße 27B, 07745 Jena, Germany.
authors: Jana
authors: J
authors: Becher
authors: INNOVENT e. V., Biomaterials Department, Pruessingstraße 27B, 07745 Jena, Germany.
authors: Stephanie
authors: S
authors: Möller
authors: INNOVENT e. V., Biomaterials Department, Pruessingstraße 27B, 07745 Jena, Germany.
authors: Matthias
authors: 0000-0002-1968-8058
authors: M
authors: Schnabelrauch
mesh_terms: D065966:Absorption, Physicochemical; D000818:Animals; D002417:Cattle; D006820:Hyaluronic Acid / Q000737:chemistry / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008970:Molecular Weight; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet; D013431:Sulfates / Q000378:metabolism*; D013737:Testis / Q000201:enzymology*
chemical_list: D013431:Sulfates; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase