- 學位論文
牛蛙輸卵管中膠原蛋白之萃取與應用
Extraction and application of collagen from bullfrog (Rana catesbeiana) fallopian tubes
摘要
膠原蛋白(collagen)為動物體中含量最豐富的蛋白質，約佔總體蛋白質的30%，廣泛應用於食品、化妝品及生醫材料上。目前已發現九大類共二十七型，每一型膠原蛋白在結締組織中皆具有不同的功能，其中以第一型含量最豐富，約佔總體膠原蛋白90%以上。但膠原蛋白來源仍多限制於哺乳類動物（牛、豬皮）、家禽類（雞）。屏東縣為全台最大牛蛙(bullfrog)產地，在肉品加工過程中會產生許多廢棄物被丟棄，例如：輸卵管(fallopian tube)。文獻指出蛙類輸卵管中含有膠原蛋白，故本研究以牛蛙輸卵管為材料，利用含胃蛋白酶(pepsin)的0.5 M醋酸於4℃下萃取48小時，並分析膠原蛋白之物化特性。由結果得知膠原蛋白萃取率約為16.35%。在SDS-PAGE電泳圖上顯示牛蛙輸卵管膠原蛋白是由兩條不同之α chains (α1和α2)所組成，其分子量分別為138 kDa及115 kDa，為第一型膠原蛋白。為提高產品之附加價值，進一步將萃取出之膠原蛋白應用於膠原蛋白保溼面膜(mask)的製造上，並測定其安定性可維持三個月以上，含0.5%、1.0%及2.0%膠原蛋白的保溼面膜之pH值約為5.25~5.73，與人體正常皮膚相似，在安全性試驗上不會有導致人體皮膚紅腫的現象產生，在有效性上使用後能增加皮膚的保水性及彈性。
並列摘要
Collagen is the most abundant protein of animal origin, comprising approximately 30% of total protein in most animals, it has been utilized in food, cosmetics and biomedical materials. There are least 27 variants of collagen, and each collagen type has different functions. TypeⅠcollagen is the major protein, comprising approximately 90% of total collagen. The main source of industrial collagen are limited to the mammals (ex. bovine and pig), bird (ex. chicken). Bullfrog is an important commercial meat in Taiwan, especially in Pingtung country. At the same time, a larger amount of fallopian tubes would be dumped as processing waste. The fallopian tubes of bullfrogs contain collagen; therefore, the aim of this study was to investigate the extraction and characterization of collagen from the fallopian tubes of bullfrogs. Collagen was digested with pepsin in 0.5 M acetic acid solution. The extraction was performed at a temperature of 4℃ for 48 hours. The result showed that the extraction yield of collagen was about 16.35%. SDS-PAGE showed that the collagen from the fallopian tubes of bullfrogs contained two different chains (α1 and α2), the molecular weight were 138 kDa and 115 kDa, and were characterized to be typeⅠ. In order to evaluate the additional value of products, we applied collagen to the collagen moisture mask manufacture. The collagen moisture masks containing 0.5%, 1.0% and 2.0% collagen have a pH value between 5.25~5.73 similar to the pH of normal human skin. The stability of collagen moisture mask was stable for more than 3 months. In safety experiment, there have no erythema was ovservated based on the Daize score test. It was also observed that the skin applied with mask containing collagen enhances the water holding capacity and elasticity of the skin.