Extraction Process Optimization and Quality Verification of Undenatured Type Ⅱ Collagen
The fresh chicken sternum cartilage was used as raw material,which was washed and sterilized followed by being broken.After being treated by neutral salt and alkali,pepsin was added into the suspension for extracting undenatuted type Ⅱ collagen by acidic and enzymetic methods.The pH and concentration of salting-out were determined by sodium lauryl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).The optimum conditions were determined by the change of the density of light bands during the extraction process.At last,the freeze-dried products of collagen obtained by salting-out at different pH and optimum salting-out concentration were analyzed by fourier transform infrared spectroscopy(FTIR),circular dichroism spectrum(CD),ultraviolet spectrophotometer(UV),hydroxyproline,isoelectric point,melting temperature(Tm),thermal decomposition temperature(Td)sodium lauryl,sulfate-polyacrylamide gel electrophoresis,amino acid content,scanning electron microscope(SEM)and atomic force microscopy(AFM).The results showed that the natural fibers of type Ⅱ collagen obtained under pH13,enzymolysis for 48 h and salting-out concentration of 0.75 mol/L exhibited a complete structure and the yield was 68.9％,the content of hydroxyproline was 11.04％.
chicken sternum cartilageundenatured type Ⅱ collagen proteinextraction qualitycharacterization analysis
万方数据
维普
