Department of Microbiology, Osmania University, Hyderabad, India.
Bioprocess Biosyst Eng. 2011 May;34(4):403-9. doi: 10.1007/s00449-010-0483-x. Epub 2010 Nov 16.
A serine alkaline protease from a newly isolated alkaliphilic Bacillus altitudinis GVC11 was purified and characterized. The enzyme was purified to homogeneity by acetone precipitation, DEAE-cellulose anion exchange chromatography with 7.03-fold increase in specific activity and 15.25% recovery. The molecular weight of alkaline protease was estimated to be 28 kDa by SDS PAGE and activity was further assessed by zymogram analysis. The enzyme was highly active over a wide range of pH 8.5 to 12.5 with an optimum pH of 9.5. The optimum temperature of purified enzyme was 45 °C and Ca(2+) further increased the thermal stability of the enzyme. The enzyme activity was enhanced by Ca(2+) and Mg(2+) and inhibited by Hg(2+). The present study is the first report to examine and describe production of highly alkaline protease from Bacillus altitudinis and also its remarkable dehairing ability of goat hide in 18 h without disturbing the collagen and hair integrity.
一种从新分离的嗜碱芽孢杆菌 GVC11 中提取的丝氨酸碱性蛋白酶被纯化并进行了特性分析。该酶通过丙酮沉淀、DEAE-纤维素阴离子交换层析进行了纯化，比活提高了 7.03 倍，回收率为 15.25%。SDS-PAGE 分析表明，碱性蛋白酶的分子量约为 28 kDa，通过酶谱分析进一步评估了其活性。该酶在 pH8.5 到 12.5 的较宽范围内具有高度活性，最适 pH 为 9.5。纯化酶的最适温度为 45°C，Ca(2+)进一步提高了酶的热稳定性。该酶的活性受 Ca(2+)和 Mg(2+)增强，受 Hg(2+)抑制。本研究首次报道了从嗜碱芽孢杆菌中生产高碱性蛋白酶的情况，并且该酶在 18 小时内对山羊皮具有显著的脱毛能力，而不会破坏胶原蛋白和毛发的完整性。
