摘要
以新鲜鸡胸软骨作为原材料,清洗灭菌后进行破碎处理,经中性盐及碱共作用的方法进行预处理后,使用酸法及酶法相结合的提取方法添加胃蛋白酶提取非变性Ⅱ型胶原蛋白。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法测定不同盐析pH值及浓度条件,和盐碱处理后材料酶解过程不同时间段光条带密度变化以确定最适条件。最后对不同盐析pH值、最佳盐析浓度条件下所获得胶原进行傅里叶红外吸收图谱,圆二色谱,紫外光谱,羟脯氨酸含量,等电点,熔融温度(T_(m)),热分解温度(T_(d)),十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,氨基酸含量,扫描电子显微镜及原子力显微镜对比分析。结果表明在pH13的混合清洗液,酶解48 h及酸性条件0.75 mol/L盐析浓度条件下所获得的Ⅱ型胶原冻干产品天然纤维结构完整,产率高达68.9%,羟脯氨酸含量可达11.04%。
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(T_(m)),thermal decomposition temperature(T_(d))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%.
作者
魏永泽
周娟
陈敬华
WEI Yongze;ZHOU Juan;CHEN Jinghua(School of Life Sciences and Health Engineering,Jiangnan University,Wuxi 214122,China)
出处
《食品科技》
CAS
北大核心
2024年第6期194-203,共10页
Food Science and Technology
基金
国家重点研发计划科技计划项目(2021YFC2103102)。
关键词
鸡胸软骨
非变性Ⅱ型胶原蛋白
提取质量
表征分析
chicken sternum cartilage
undenatured type Ill collagen protein
extraction quality
characterization analysis
