Kakkar Prachi, Madhan Balaraman, Shanmugam Ganesh
Central Leather Research Institute, Council of Scientific and Industrial Research, Adyar, Chennai, 600020 India.
Springerplus. 2014 Oct 10;3:596. doi: 10.1186/2193-1801-3-596. eCollection 2014.
Keratin from the hoof is a less explored source for making valuable products. In this paper we present the extraction of pure keratin from bovine hooves and characterized them to better address the possible exploitation of this bio-resource as an alternative material for tissue engineering applications. The keratin protein from the pulverized hooves was extracted by reduction, which was observed to be pure, and two polypeptide chains of molecular weight in the range of 45-50 and 55-60 KDa were determined using SDS-PAGE assay. FTIR analysis complementing circular dichroism (CD) data, established that hoof keratin predominantly adopted α-helical conformation with admixture of β-sheet. The keratin was shown to have appreciably high denaturation temperature (215°C) as indicated by differential scanning calorimetric (DSC) analysis. Thermogravimetric analysis (TGA) also showed the retention of 50% of the original weight of the sample even at a temperature of 346°C. The keratin from the hoof had been observed to be biocompatible when analyzed with MTT assay using fibroblast cells, showing more than 90% cell viability. Hence, hoof keratin would be useful for high value biomedical applications.
蹄角蛋白是一种尚未得到充分探索的用于制造有价值产品的原料。在本文中,我们展示了从牛蹄中提取纯角蛋白并对其进行表征,以更好地探讨将这种生物资源用作组织工程应用替代材料的可能性。通过还原法从粉碎的蹄中提取角蛋白,经观察其为纯品,使用SDS - PAGE分析确定了分子量在45 - 50 kDa和55 - 60 kDa范围内的两条多肽链。傅里叶变换红外光谱(FTIR)分析结合圆二色性(CD)数据表明,蹄角蛋白主要采用α - 螺旋构象,并伴有β - 折叠。差示扫描量热法(DSC)分析表明,角蛋白具有相当高的变性温度(215°C)。热重分析(TGA)也显示,即使在346°C的温度下,样品仍保留其原始重量的50%。在用成纤维细胞进行MTT分析时,观察到蹄角蛋白具有生物相容性,细胞活力超过90%。因此,蹄角蛋白将有助于高价值生物医学应用。