Extraction，purification And Identification Of Collagen From Bovine Lung Tube

Collagen not only have some special functions but contains rich nutrition,so it is a kind of high value product. It exists extensively,especially found in animal connective tissues. When we slaughtered the Livestock and poultry a large number of by-products have to come into existence and have not been exploited to result in waste. Livestock and poultry bones are a kind of natural resources, collagen is one of them,which have abundant with nutrition. Therefore, It is necessary to further investigates the by-products in the process until they can be used. It has the significant diretive meaning to widen collagen extion resources and application.In this paper, looking for a new raw materials to extracting collagen products-the bovine lung tube. Determination of nutritional components in bovine lung tube:Moisture content, protein content, fat content, mineral content and collagen content, the results showed that:the content of collagen,18.81%±0.12, ash content1.57%, water content66.89%, crude protein content of20.83%, fat content of9.42%mineral elements, Ca1197.27mg/Kg, Fe46.98mg/Kg, Mg341.90mg/Kg, Al15.11mg/Kg,Cu0.75mg/Kg, Zn8.16mg/Kg, Pb3.26mg/Kg. Experiment results demonstrate that in bovine lung tube contains a lot of collagen,content of18.81%±0.12.Application of different enzymes to extract collagen from bovine lung tubc. According to cross experiment and response surface design to find out the types of the enzymes and the best extracting conditions:the pH of Alkaline proteinase is8.70、solid-liquid ratio is7%、enzyme addition levels(E/S)is2.0%、 enzymolysis temperature is34.86℃、action time is24.28h. In order to obtain high-purity, high extraction rate, before extraction of collagen by Alkaline proteinase we should do the following: degrease、removing proteins、 de-calcification、 cartilage was defatted with6%sodium carbonate degreased,using solid-liquid ratio is1:20soak12h and then using2.0mol/L NaCl extracted to remove contaminating protein. Selected EDTA solution to De-calcification, Based on orthogonal experiment educe what conditions get the best:use0.20mol/L EDTA、action time is36h、 solid-liquid ratio is1:20. Decalcification rate is91.73%.DEAE-52chromatography provided a rapid and efficient method for separating collagen from proteoglycans in cartilage extracts. The results of SDS-PAGE and UV spectra suggested the prepared type II collagen the prepared type II collagen essentially was free from contaminating proteins by the method used in this study for purification. The extracted collagen was identified as type II collagen and the purity reached electrophoretic grade.At the same time the results show that collagen exhibited a maximum absorbance at227nm, but little absorbance near250-290nm.This experiment analyzes the amino acid components of collagen, Make a comparative amino acid components between collagen from bovine lung tube and the type Ⅱ collagen produced by Sigma Company,We found they resemble each other. To sum up, This prepared collagen of type Ⅱ is a product of high purity which has great medical value. this study, sought out another new resources to extraction type Ⅱ collagen. At the same time, the results of this thesis might be some useful to extracting collagen.