Muindi Munguti Peter, Lee Ji Hae, Kweon HaeYong, Kasina Muo
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.
Polymers (Basel). 2022 Oct 1;14(19):4118. doi: 10.3390/polym14194118.
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.
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。