暗纹东方鲀鱼皮胶原蛋白的提取及其特性

为增加暗纹东方鲀附加值,考察不同处理方法对提取暗纹东方鲀鱼皮胶原蛋白的影响,实验以暗纹东方鲀鱼皮为对象,通过热水法、酸法和胃蛋白酶、木瓜蛋白酶、无花果蛋白酶处理鱼皮提取胶原蛋白。通过对不同胶原蛋白的提取率、氨基酸组成、傅里叶红外光谱(FTIR)、紫外光谱(UV)、扫描电镜、十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)进行研究,对暗纹东方鲀鱼皮胶原蛋白特性进行表征和比较。结果显示,用胃蛋白酶得到的胶原蛋白提取率最高;氨基酸组成相似但含量不同;3种酶制备的胶原蛋白中,脯氨酸含量显著低于酸和热水制备的胶原蛋白;FTIR扫描结果表明,5种处理方法得到的胶原蛋白都存在Amide A、Amide B、AmideⅠ、AmideⅡ和AmideⅢ,均保持了胶原蛋白三螺旋结构;紫外光谱显示在235 nm左右有强吸收峰,结合FTIR确定其为典型的胶原蛋白,经过SDS-PAGE分析,确定暗纹东方鲀鱼皮胶原蛋白为Ⅰ型胶原蛋白。酸法和胃蛋白酶较好地保留了胶原蛋白的β、α1和α2链,木瓜蛋白酶作用化学键比其他酶广泛,得到小分子量的胶原蛋白分子;扫描电镜结果显示,酸法提取的胶原蛋白最适合应用在生物医学材料上运载药物。由此可见,不同处理方法提取的胶原蛋白理化特性存在一定差异,不同的酶制备的胶原蛋白分子量分布会产生明显差别,可根据研究需要选用不同处理方法开发胶原蛋白产品。

Extraction and characteristics of collagens from the skin of puffer fish (Tetrodontiformes fasciatus)

In order to enhance the added value of Tetrodontiformes fasciatus and investigate the effects of different preparations to the extracted collagen from skin of T.fasciatus, collagens extracted from the skin of T. fasciatus by acid, hot water, pepsin, papain, and ficin were used to evaluate chemical and structural properties, respectively. In this study, characterization of collagen from T.fasciatus skin in term of the extraction yield, amino acid composition, Fourier transform infrared (FTIR) spectroscopy, ultraviolet spectroscopy(UV), scanning electron microscopy(SEM) and sodium dodecyl sulfate-polyacrylamide gel electropheresi (SDS-PAGE). Results showed that the extraction yield of collagen obtained by pepsin was the highest,the amino acid compositions were similar and the contents were different,the content of proline in collagen extracted by 3 enzymes is significantly lower than that of collagen extracted by acid and hot water,FTIR showed collagen obtained by the five extraction methods had Amide A, Amide B, Amide I, Amide II and III, and confirmed the triple helical structure of the collagens,UV showed a strong absorption peak around 235 nm, indicating a typical collagen when combined with FTIR results,SDS -PAGE analysis confirmed that collagen in skin of T. fasciatus was type I collagen, and preparations by acid and pepsin retained protein structure while papain dissociated collagen and obtained protein with low molecular mass,SEM showed that extracted collagen by acid was most suitable for biomedical materials. Therefore, different extraction methods resulted in different chemical and structural properties of ?sh skin collagens. The molecular weight distribution of collagen extraction by different enzymes had obvious differences. According to research needs, different preparations are selected to develop collagen products.