Purification and Characterization of Manganese-Dependent Alkaline Serine Protease from Bacillus pumilus TMS55
Purification and Characterization of Manganese-Dependent Alkaline Serine Protease from Bacillus pumilus TMS55
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DOI：
10.4014/jmb.1009.09001
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发表时间：
2011-01-01
影响因子：
2.8
通讯作者：
Pandian, Shunmugiah Karutha
中科院分区：
文献类型：
--
作者：
Ibrahim, Kalibulla Syed;Muniyandi, Jeyaraj;Pandian, Shunmugiah Karutha
研究了短小芽孢杆菌TMS 55产生的一种锰离子依赖型碱性丝氨酸蛋白酶的纯化及性质。该酶的纯化分三步进行：用硫酸铵沉淀浓缩粗酶，然后通过凝胶过滤和阳离子交换层析。纯化的蛋白酶具有约35 kDa的分子量，在7.0至12.0的宽pH范围内具有高活性，并且在7.5至11.5的pH范围内保持稳定。发现酶活性的最适温度为60 ℃。PMSF和AEBSF（1 mM）显著抑制蛋白酶活性，表明该蛋白酶是丝氨酸蛋白酶。Mn ~（2+）离子增强了酶的活性和稳定性。此外，纯化的蛋白酶与氧化剂（H2 O2，2%）和有机溶剂（25%），如苯，己烷和甲苯保持稳定。因此，该蛋白酶的这些特性及其脱毛能力表明其具有广泛的商业应用潜力。
The purification and characterization of a Mn2+-dependent alkaline serine protease produced by Bacillus pumilus TMS55 were investigated. The enzyme was purified in three steps: concentrating the crude enzyme using ammonium sulfate precipitation, followed by gel filtration and cation-exchange chromatography. The purified protease had a molecular mass of approximately 35 kDa, was highly active over a broad pH range of 7.0 to 12.0, and remained stable over a pH range of 7.5 to 11.5. The optimum temperature for the enzyme activity was found to be 60 degrees C. PMSF and AEBSF (1 mM) significantly inhibited the protease activity, indicating that the protease is a serine protease. Mn2+ ions enhanced the activity and stability of the enzyme. In addition, the purified protease remained stable with oxidants (H2O2, 2%) and organic solvents (25%), such as benzene, hexane, and toluene. Therefore, these characteristics of the protease and its dehairing ability indicate its potential for a wide range of commercial applications.