Studies On The Extraction, Structure And Vitro Activity Of Collagen And Dermatan Sulfate From Lophius Litulon Skin

The extraction, structure and activity of collagen and dermatan sulfate (DS) from the skin of Halieutaea stellata was investigated in this paper. The research is to provide a theoretical basis for utilization of Halieutaea stellata skin and development of collagen.1. The collagen and DS was obtained from the skin of Halieutaea stellata. Collagen and DS was extracted under the following:papain dosage was4000IU/g, solid to liquid ratio was1:20, the pH was adjusted to7, at50℃reaction for6h., and the maximum extraction yield of collagen was59.4%, the DS was52.9mg/100g.2. The physicochemical charateristics of Halieutaea stellata skin collagen was studied. SDS-PAGE suggested that collagen was comprised of a1, a2, and3chain, might be classified as type I collagen. Amino acid analysis showed that the content of histidine was the highest, reaching22.53%, the amino acid composition showed that this collagen was type Ⅰ.The denaturation temperature of collagen was22.3℃by viscosity determined. It was lower than those from pigskin collagen28.4℃ and donkey-hide gelatin29.2℃.The results suggested that thermal stability of collagens were highly depending on animal species, growth environment and so on. The maximum ultraviolet absorption of collagen was227nm, which was contributed by n-π*transition of C=O in the peptide bond.3. Chemical methods were used to analysis the basic constituents of DS. It showed that the DS from Halieutaea stellata skin contains hexosamine(31.2%), uronic acid(30.46%), moisture content(7.63%) and N content(2.35%). Electrophoresis analysis showed that the molecular weight was15-22kDa, whose result demonstrated that this DS without other glycosaminoglycans. Disaccharide analysis showed that four kinds of disaccharide with different number and substituent sites of SO3were identified. Propotion of each disaccharide (accounting for the proportion of total disaccharide) were△Di-OS (18.5%),△Di-6S (30.1%), ADi-4S (44.9%),△Di-diSB (6.5%). Then, methods for analyzing and identifying products derived from partially enzymatic degradation of DS by HPLC-ESI-MS(MS/MS) were used. After ESI-MS analysis and structural characterization of mixtures of DS-derived oligosaccharides, structures of these fragments were initially identified as follows:dp2:IdoAS1-GalNAcS1;dp4:dIdo AS1-GalNAcS1-IdoAS1-GalNA;dp6:dldoAS1-GalNAcS1-IdoAS1-GalNAcS1-IdoA-GalNAcS1; dp8:dIdo AS1-GalNAcS1-IdoAS1-GalNAcS1-IdoA-GalNAcS1-IdoA-GalNAcS1;dp10:dIdoA-GalNAc-(IdoA-GalNAc)4。4. The hygroscopicity and moisture retention of the collagen from Halieutaea stellata skin was studied. The collagen showed very good moisture absorption and moisture resistance. So it can be used as cosmetic products and raw materials. In addition, the anticoagulant activity of the DS from Halieutaea stellata skin was invested with heparinum and other GAGs as control. The results showed that the DS has significant effect on anticoagulant activity which indicating that the DS mainly effect on intrinsic pathway.5. Vitro osteoclast inhibition tests showed DS can significantly inhibit the growth of osteoclasts, whose inhibition rate reached57%, which has a potential value as drug development to alleviate osteoporosis and rheumatoid arthritis.