title: Characterisation of separated end hyaluronan oligosaccharides from leech hyaluronidase and evaluation of angiogenesis.
abstract: Hyaluronan oligosaccharides (o-HAs), especially saturated o-HAs, have attracted intensive attention due to their potential applications in medical treatments. In this study, the hydrolysis process of leech hyaluronidase (LHase) towards the hyaluronan was investigated by HPLC and HPLC/ESI-MS. The proportions of hyaluronan tetrasaccharide (HA4) with hexasaccharide (HA6), end products, were illustrated to have a relationship with the amount of LHase. Higher yield of HA4 was achieved with higher activity of LHase. After optimisation of the packing resin and operation parameters (balanced pH, elution concentration, elution volume and elution flow rate), the highly pure HA4 and HA6 were efficiently separated and prepared by combining ion exchange Q-Sepharose Fast Flow and size exclusion column chromatography. Compared with o-HAs (average Mr of 4000 Da), HA4 and HA6 were demonstrated to show higher activity for promoting angiogenesis, which was similar with the corresponding HA4 and HA6 produced by bovine testicular hyaluronidase. The pure HA4 and HA6 that prepared from LHase will attract intensive studies and be used in potential applications in near future.
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, Jiangsu, China.
authors.forename: Mengxian
authors.initials: M
authors.lastname: Lv
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, Jiangsu, China. Electronic address: mwang423@126.com.
authors.forename: Miao
authors.initials: M
authors.lastname: Wang
authors.affiliation: Laboratory of Tumor Pharmacology, Wuxi Medical School, Jiangnan University, Wuxi 214122, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Cai
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, Jiangsu, China.
authors.forename: Wenxing
authors.initials: W
authors.lastname: Hao
authors.affiliation: The Key Laboratory of Industrial Biotechnology, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Panhong
authors.initials: P
authors.lastname: Yuan
authors.affiliation: The Key Laboratory of Industrial Biotechnology, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Kang
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006820:Hyaluronic Acid / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D006868:Hydrolysis; D007865:Leeches / Q000201:enzymology; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D043925:Angiogenesis Inducing Agents; D009844:Oligosaccharides; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase
keywords: Angiogenesis; Chromatography; Hyaluronan hexasaccharide; Hyaluronan tetrasaccharide; Leech hyaluronidase; Quantitative analysis