【摘要】 【目的】分析不同提取方法对龟甲胶原蛋白得率、纯度、结构特征和理化特性的影响。【方法】采用超声联合酶法、酸法、酶法提取龟甲胶原蛋白。【结果】超声联合酶法提取的龟甲胶原蛋白的得率和纯度分别为2.29%和85.65%，显著高于酸法（0.78%、73.15%）和酶法（1.77%、75.63%）。紫外吸收光谱、红外吸收光谱、圆二色谱和电泳图谱显示，3种方法提取的龟甲胶原蛋白均为Ⅰ型胶原蛋白，具有较完整的三螺旋结构。X射线衍射图谱表明，超声联合酶法提取的龟甲胶原蛋白的分子结构更为紧凑、有序度更高。超声联合酶法和酶法提取的龟甲胶原蛋白的粒径均分布在340 nm左右，但超声联合酶法提取的龟甲胶原蛋白的粒径分布集中度更低。3种方法提取的龟甲胶原蛋白的热变性温度（T_d）均为44℃，扫描电子显微镜结果显示，超声联合酶法提取的龟甲胶原蛋白的微观结构更加均匀、规则。【结论】该研究利用的超声联合酶法为龟甲胶原蛋白的提取提供了新思路。与常规的酸法和酶法相比，超声联合酶法提取的龟甲胶原蛋白的得率和纯度均较高，表明该方法具有广阔的应用前景。更多还原

【Abstract】 [Objective] The effects of different extraction methods on the yield, purity, structural characteristics, and physicochemical properties of tortoise shell collagen were investigated. [Method] The ultrasound-assisted enzymatic method, acid method, and enzymatic method were used to extract collagen from tortoise shell powder. [Result] The yield and purity of tortoise shell collagen extracted by ultrasound-assisted enzymatic method were 2.29% and 85.65%, respectively, significantly higher than those extracted by acid method(0.78% and 73.15%) and enzymatic method(1.77% and 75.63%). Ultraviolet absorption spectroscopy, infrared absorption spectroscopy, circular dichroism spectroscopy, and electrophoresis profiles showed that collagens extracted from tortoise shell by three methods were all type I collagen and retained intact triple helix structures. X-ray diffraction patterns revealed that the molecular structure of tortoise shell collagen treated with ultrasound-assisted enzymatic method was more compact, with a higher degree of order. The particle sizes of tortoise shell collagens extracted by ultrasound-assisted enzymatic method and enzymatic method were both distributed around 340 nm, while tortoise shell collagen extracted by ultrasound-assisted enzymatic method exhibited a lower particle size distribution. The thermal denaturation temperatures(T_d) of the tortoise shell collagens extracted by the three methods were all 44 ℃. Scanning electron microscopy results showed that the microstructure of tortoise shell collagen extracted by ultrasound-assisted enzymatic method was more uniform and regular. [Conclusion] The combination of ultrasound treatment and enzymatic methods provide a new perspective for the extraction of collagen from tortoise shell. Compared with the conventional acid method and enzymatic method, the ultrasoundassisted enzymatic extraction method shows improvements in the yield and purity of tortoise shell collagen, thus demonstrating a broad application prospect.更多还原