以暗纹东方鲀和红鳍东方鲀鱼皮为原料,分析比较其胶原蛋白的理化及体外抗氧化特性。采用胃蛋白酶对胶原蛋白进行提取,分析氨基酸组成,采用紫外和红外光谱、十二烷基硫酸钠聚丙烯酰胺凝胶电泳、扫描电镜、原子力显微镜等技术对2种鱼皮胶原蛋白的理化特性进行比较,采用DPPH自由基及·OH的清除率等指标对其体外抗氧化活性进行分析。结果表明,2种鱼皮胶原蛋白氨基酸含量差异不显著(P>0.05);光谱扫描结果显示,2种胶原蛋白均为三螺旋结构完整的I型胶原蛋白;2种胶原蛋白的氨基酸组成和蛋白质二级结构呈显著相关性;此外,暗纹东方鲀鱼皮胶原蛋白(Takifugu obscurus skin collagen,TOSK)的热变性温度为37 ℃,红鳍东方鲀胶原蛋白(Takifugu rubripes skin collagen,TRSK)的热变性温度为35 ℃,微观结构观察显示TOSK和TRSK都具有较为完整的纤维网状结构,且TOSK丝状结构比TRSK更光滑。0.8 mg/mL的TOSK对DPPH自由基及·OH的清除率分别为45.75%及63.57%;0.8 mg/mL TRSK对DPPH自由基的清除率为44.94%,在1.0 mg/mL时对·OH的清除率为63.54%。与市售鱼皮胶原蛋白相比,提取的2种河豚鱼皮胶原蛋白的抗氧化性能优于市售鱼皮胶原蛋白,该研究可为胶原蛋白在化妆品领域的应用提供理论基础。
In this study, the skins of Takifugu obscurus and Takifugu rubripes were used as raw materials to analyze and compare the physicochemical and in vitro antioxidant properties of their collagen. The collagen was extracted by pepsin, and the physicochemical properties of the two fish skin collagens were compared by analyzing the amino acid composition, ultraviolet and infrared spectra, sodium dodecyl sulfate polyacrylamide gel electrophoresis, scanning electron microscopy, and atomic force microscopy. The antioxidant activity in vitro was analyzed by the scavenging rate of DPPH and ·OH free radicals. Results showed that there was no significant difference in amino acid contents between the two kinds of collagen (P>0.05) and the spectral scanning results showed that both collagens were type I collagen with a complete triple helix structure. The two collagens correlated significantly in amino acid composition and protein secondary structure. In addition, the thermal denaturation temperature of T. obscurus skin collagen (TOSK) was 37 ℃, and that of T. rubripes skin collagen (TRSK) was 35 ℃. The microstructural observation showed that both TOSK and TRSK had a relatively complete fibrous network structure, and the filamentous structure of TOSK was smoother than that of TRSK. The scavenging rates of TOSK on DPPH and ·OH free radicals were 45.75% and 63.57% at 0.8 mg/mL respectively, while the scavenging rates of TRSK on DPPH and ·OH free radicals were 44.94% and 63.54% at 0.8 mg/mL and 1.0 mg/mL, respectively. The antioxidant effect of the extracted puffer fish skin collagen was better than that of commercial fish skin collagen. This study could provide a theoretical basis for the application of collagen in cosmetics.
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