The study of silky fowl feet enzymatic hydrolysates onproperty of in vitro antioxidative and cell photo-protection

• Main Authors: Pei-Chieh Tsai, 蔡佩潔
• Other Authors: 林亮全
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/55921511062218665786

碩士 === 國立中興大學 === 動物科學系所 === 97 === Solar ultraviolet (UV) radiation can trigger certain signals and effects in skin cells, causing oxidative stress resulting in pathologies. Exposure of cells to UV has been shown to alter the activity of various proteins in the plasma membrane, cytoplasm and nucleus of the cells, which indirectly caused the aged on appearance of skin. Recently, there are lots of physiological and biochemistry researches indicated that proteins not only provided the amino acid, but also produced the bioactive peptides which have kinds of functional properties after hydrolyzed by enzymes. Silky fowl feet contained meat, cartilage, bone marrow and melanin (in skin and periosteum), Thus, the aim of this study was to nvestigate the functional characteristics of enzymatic hydrolysates obtained from silky fowl feet. Alcalalse(A), Flavorzyme(F) and Protase N(P) were used for two-stage hydrolysis respectively. Two kinds of enzymes were added in turn for the preparation of one sample, and there were six combinations (AF, AP, FA, FP, PA and PF). After hydrolysis for 4 hours, the hydrolysates were be lyophilized and analyzed the electrophoresis, soluble protein, peptide content, UV absorption spectrum, melanin content, yrosinase activity inhibition and antioxidative activities. Besides, human skin fibroblast (WS1) was cultured with silky fowl feet hydrolysates and nvestigated the cell viabilities induced by hydrogen peroxide, tert-butyl hydroperoxide (t-BHP) and UV light. The enzyme activities of catalase, superoxide dismutase, glutathione peroxidase and the content of glutathione and collagen of WS1 were determined. The results showed that the soluble protein were decreased and peptide content of the hydrolysates were increased respectively, which indicated the well hydrolysis in current study. All hydrolysates had broad spectrum absorption and tyrosinase inhibitory activity. Especially, PA had the highest absorption and tyrosinase inhibitory ability. Melanin content analysis showed silky fowl feet hydrolysates contained melanin range from 1.54 to 2.32 μg/mg. In the in vitro antioxidative properties, all the silky fowl feet hydrolysates exhibited effects on inhibition of the lipid peroxidation, radical DPPH scavenging effects, reducing ability and ferrous ion chelating ability. In cell cultures of WS1, all hydrolysates exhibited free radical scavenging ability which could protect cells from death induced by hydrogen peroxide and t-BHP. The cell viabilies of WS1 treated with silky fowl feet hydrolysates increased dramatically and preserved the whole appearances of cell without damages, afetr UV irradiation. In particular, PA had the best activity. Moreover, increasing expression of catalase, superoxide dismutase and glutathione peroxidase activity were observed in cells treated with hydrolysates. And glutathione and collagen content increased in hydrolysates treatments. The results in this study suggested that silky fowl feet hydrolysates could protect cell from damage by effects on scavenging free radicals, and possess high potential to be utilized as naturally photo-protection ingredient of cosmetic bleaching products which can promote activities of molecular antioxidant enzymes activity in fibroblast.