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提取成分对丝胶蛋白(SS)构象和稳定性的影响。

Effect of Extraction Ingredients on the Conformation and Stability of Silk Sericin (SS).

作者信息

Muindi Munguti Peter, Lee Ji Hae, Kweon HaeYong, Kasina Muo

机构信息

National Sericulture Research Center, Kenya Agricultural and Livestock Research Organization, Thika 7816-01000, Kenya.

National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Korea.

出版信息

Polymers (Basel). 2022 Oct 1;14(19):4118. doi: 10.3390/polym14194118.

Abstract

Silk sericin (SS) has different physicochemical properties depending on the extraction technique. In this study, SS was isolated in the presence of ingredients, including 5 to 10% ethanol (EtOH) and 5 to 10% glycine. Furthermore, temperature conditions of 80 °C, 100 °C, and 120 °C were used for 1, 3, and 5 h to evaluate the extraction rates. The extraction, gelation, structural, and cytotoxicity properties of SS extracted under different conditions were investigated. Extraction at 100 °C and 120 °C were found to have the highest SS yield, with 80 °C being the lowest. SS isolated at 100 °C and 120 °C for 1 and 3 h in water, and EtOH gelled at 4 °C in 2 to 3 days and 37 °C in 40 min. Glycine SS extracts were obtained at 100 °C and 120 °C for 1 h, gelled at 4 °C for 20 days and 37 °C for 16 h. SS was observed at 80 °C, with no gelation occurring. Glycine SS extracts obtained for 3, and 5 h at 120 °C showed no gelation. Circular dichroism (CD) results show glycine in SS induces α-helix and random coil structure. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and fast performance liquid chromatography (FPLC) were used to quantify the molecular weight distribution at 63 and 70 kDa, respectively. The MMT assay (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) revealed no cytotoxicity in macrophage RAW 264.7 cells treated with this method SS; these findings present the significance and possibility of using selected extraction ingredients in SS that allow for the application of native SS at an initial extraction viscosity.

摘要

丝胶蛋白(SS)因提取技术的不同而具有不同的物理化学性质。在本研究中,丝胶蛋白是在含有5%至10%乙醇(EtOH)和5%至10%甘氨酸的成分存在下分离得到的。此外,分别在80℃、100℃和120℃的温度条件下提取1小时、3小时和5小时,以评估提取率。研究了在不同条件下提取的丝胶蛋白的提取、凝胶化、结构和细胞毒性特性。结果发现,在100℃和120℃下提取的丝胶蛋白产率最高,80℃下提取的产率最低。在100℃和120℃下于水中提取1小时和3小时得到的丝胶蛋白,在乙醇中于4℃下2至3天凝胶化,在37℃下40分钟凝胶化。在100℃和120℃下提取1小时得到甘氨酸丝胶蛋白提取物,在4℃下20天凝胶化,在37℃下16小时凝胶化。在80℃下观察到丝胶蛋白,但未发生凝胶化。在120℃下提取3小时和5小时得到的甘氨酸丝胶蛋白提取物未出现凝胶化。圆二色性(CD)结果表明,丝胶蛋白中的甘氨酸诱导α-螺旋和无规卷曲结构。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和快速高效液相色谱(FPLC)分别用于定量63 kDa和70 kDa处的分子量分布。MTT法(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)显示,用该方法处理的丝胶蛋白对巨噬细胞RAW 264.7细胞无细胞毒性;这些发现表明了在丝胶蛋白中使用选定提取成分的意义和可能性,这使得天然丝胶蛋白能够在初始提取粘度下得到应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d47c/9572401/7d6a95c43324/polymers-14-04118-g001.jpg

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