该研究采用深共熔溶剂(deep eutectic solvents,DESs)提取金枪鱼皮胶原蛋白(tuna skin collagen,TSCO),首先以TSCO提取率及高级结构的完整度为评价指标选择提取效果较好的溶剂组合,随后讨论不同提取条件的影响,包括组分比例、提取转速、温度和时间,最后通过柱层析法纯化粗提TSCO。利用傅里叶变换红外光谱、SDS-PAGE和扫描电子显微镜等技术对提取的胶原蛋白进行分析。结果表明,在甜菜碱、柠檬酸、水物质的量比2∶1∶30,提取温度60 ℃,转速1 000 r/min,提取时间180 min的条件下,胶原蛋白的提取率可达到98.74%。经过柱层析法纯化后其纯度为91.4%,高于胶原蛋白标准品纯度(80.4%)。结构分析显示,提取的TSCO保持了完整的三螺旋结构,分子质量分布均匀并具有良好的微观形貌,说明DESs能高效提取TSCO且没有破坏其结构的完整性。该研究证明了DESs在TSCO提取中的高效性和可行性,为促进金枪鱼皮的高值化利用提供参考。
In this study, deep eutectic solvents (DESs) were used to extract tuna skin collagen (TSCO).Firstly, the TSCO extraction rate and the integrity of the advanced structure were used as evaluation indexes to select the solvent combinations with better extraction effect.Subsequently, the effects of different extraction conditions were discussed, including the proportion of components, extraction speed, temperature and time, and finally the crude extracted TSCO was purified by column chromatography.The extracted collagen was analyzed using Fourier transform infrared spectroscopy, SDS-polyacrylamide gel electrophoresis and scanning electron microscopy.The results showed that the extraction rate of collagen could reach 98.74% under the conditions of 2∶1∶30 molar ratio of betaine, citric acid and water, extraction temperature of 60 ℃, rotational speed of 1 000 r/min and extraction time of 180 min.After purification by column chromatography its purity was 91.4%, which was higher than that of the collagen standard (80.4%).Structural analysis showed that the extracted TSCO maintained an intact triple-helical structure with uniform molecular weight distribution and good microscopic morphology, indicating that DESs could efficiently extract TSCO without destroying its structural integrity.This study demonstrated the high efficiency and feasibility of DESs in TSCO extraction, which can provide a reference for promoting the high-value utilization of tuna skin.
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