Manavalan Tamilvendan, Manavalan Arulmani, Ramachandran Shiyamsundar, Heese Klaus
Centre for Advanced Studies in Botany, University of Madras, Chennai, Tamil Nadu 600 025, India.
Department of Biotechnology, Anna University, BIT-Campus, Tiruchirappalli, Tamil Nadu 620 024, India.
Biology (Basel). 2020 Dec 16;9(12):472. doi: 10.3390/biology9120472.
An increased need by the green industry for enzymes that can be exploited for eco-friendly industrial applications led us to isolate and identify a unique protease obtained from a proteolytic -TK1 strain from a seawater source. The extracellular thermostable serine protease was processed by multiple chromatography steps. The isolated protease displayed a relative molecular weight (MW) of 33 kDa (confirmed by zymography), optimal enzyme performance at pH 8.0, and maximum enzyme performance at 70 °C with 100% substrate specificity towards casein. The proteolytic action was blocked by phenylmethylsulfonyl fluoride (PMSF), a serine hydrolase inactivator. Protease performance was augmented by several bivalent metal cations. The protease tolerance was studied under stringent conditions with different industrial dispersants and found to be stable with Surf Excel, Tide, or Rin detergents. Moreover, this protease could clean blood-stained fabrics and showed dehairing activity for cow skin with significantly reduced pollution loads. Our results suggest that this serine protease is a promising additive for various eco-friendly usages in both the detergent and leather industries.
绿色产业对可用于环保工业应用的酶的需求增加，促使我们从海水来源的蛋白水解-TK1菌株中分离并鉴定出一种独特的蛋白酶。通过多个色谱步骤对细胞外热稳定丝氨酸蛋白酶进行了处理。分离出的蛋白酶相对分子量（MW）为33 kDa（经酶谱法确认），在pH 8.0时酶性能最佳，在70°C时酶性能最高，对酪蛋白具有100%的底物特异性。蛋白水解作用被丝氨酸水解酶失活剂苯甲基磺酰氟（PMSF）阻断。几种二价金属阳离子增强了蛋白酶的性能。在不同工业分散剂的严格条件下研究了蛋白酶的耐受性，发现它在汰渍、碧浪或奥妙洗涤剂中稳定。此外，这种蛋白酶可以清洁血渍织物，并对牛皮显示出脱毛活性，污染负荷显著降低。我们的结果表明，这种丝氨酸蛋白酶是洗涤剂和皮革工业中各种环保用途的有前途的添加剂。
