不同方法提取方格星虫胶原蛋白及其酶解物对HaCaT细胞的影响
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TS 254.9

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广东海洋大学大学生创新创业训练计划项目(CXXL2020004);湛江市科技发展专项资金竞争性分配项目(2019A902);近海资源生物技术福建省高校重点实验室开放性课题(2019IRB01);国家重点研发计划-蓝色粮仓(2020YFD0901104)


Different methods of extracting collagen from Sipunculus nudus and its effect of enzymatic hydrolysate on HaCaT cells
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    摘要:

    以方格星虫(Sipunculus nudus)体壁为原料,分别采用酸法、酶法和热水法提取胶原蛋白并对其理化性质进行分析;通过检测氨基酸组成、紫外光谱、傅里叶红外光谱、SEM扫描电镜分析比较了3种胶原蛋白的特性,并采用HaCaT细胞,探讨3种酶解产物对其生长活力的影响。结果表明,热水法提取胶原蛋白的提取得率高于其他两种方法,3种胶原蛋白均含有胶原特征型氨基酸甘氨酸Gly、脯氨酸Pro和羟脯氨酸Hyp。紫外和红外光谱图显示,3种胶原蛋白均在222 nm附近有明显的吸收峰,且存在酰胺A、酰胺B的吸收峰和酰胺Ⅰ、Ⅱ、Ⅲ的特征峰,提示3种胶原蛋白均属于Ⅰ型胶原蛋白,具有三股螺旋结构。而扫描电镜结果显示,相比于酶法和热水法,酸法提取的胶原蛋白三股螺旋结构保持更为完整,且较好地维持胶原纤维的原有结构。3种胶原蛋白酶解肽在50~100 μg/mL时,能够显著提高正常和UVB损伤的HaCaT细胞活力,表明方格星虫胶原蛋白的酶解产物在皮肤光老化修复方面具有较好的潜在应用前景。

    Abstract:

    The collagen was extracted by acid, enzymolysis and hot water methods respectively from the body wall of Sipunculus nudus, and their physicochemical properties were analyzed. The three collagen extracts were compared respectively by amino acid composition, UV spectroscopy, Fourier infrared spectroscopy and SEM scanning electron microscopy. The effects of the three collagen zynolytes on the growth viability of HaCaT cells were investigated. The results show that the extraction rate of collagen by hot water method is higher than the other two methods, and all three collagens contain the collagen characteristic amino acids Gly, Pro and Hyp. The UV and IR spectrograms show that all three collagens have obvious absorption peaks near 222 nm, and there are absorption peaks of amide A and amide B and characteristic peaks of amide Ⅰ, Ⅱ and Ⅲ, suggesting that all three collagens belong to type I collagen with a three-stranded helical structure. And the results of scanning electron microscopy show that the three-stranded helical structure of collagen extracted by acid method is more complete and maintains the original structure of collagen fibers better than that of enzymatic method and hot water method. The collagen peptides extracts have the ability to significantly increase the viability of normal and UVB-damaged HaCaT cells in the range of 50?100 μg/mL, indicating that the enzymatic products of collagen of Sipunculus nudus have good potential applications in skin photoaging repair.

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林海生,冯畅,王雯,袁建军,陈洪彬,杨维,曾堉斯,郑梦琪,范明珊,杨海明,邓旗.不同方法提取方格星虫胶原蛋白及其酶解物对HaCaT细胞的影响[J].上海海洋大学学报,2023,32(6):1165-1175.
LIN Haisheng, FENG Chang, WANG Wen, YUAN Jianjun, CHEN Hongbin, YANG Wei, ZENG Yusi, ZHENG Mengqi, FAN Mingshan, YANG Haiming, DENG Qi. Different methods of extracting collagen from Sipunculus nudus and its effect of enzymatic hydrolysate on HaCaT cells[J]. Journal of Shanghai Ocean University,2023,32(6):1165-1175.

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  • 收稿日期:2022-05-22
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