Decolorization of crude latex by activated charcoal, purification and physico-chemical characterization of religiosin, a milk-clotting serine protease from the latex of Ficus religiosa.
Decolorization of crude latex by activated charcoal, purification and physico-chemical characterization of religiosin, a milk-clotting serine protease from the latex of Ficus religiosa.
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DOI：
10.1021/jf101020u
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发表时间：
2010-06
影响因子：
6.1
通讯作者：
Moni Kumari;Anurag Sharma;M. Jagannadham
中科院分区：
文献类型：
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作者：
Moni Kumari;Anurag Sharma;M. Jagannadham
采用活性炭对宗教榕的粗胶乳进行脱色。脱色遵循Freundlich和Langmuir方程。用阴离子交换层析法从脱色胶乳中纯化出一种丝氨酸蛋白酶，命名为religiosin。MALDI-TOF法测得其分子量为43.4 kDa。Religiosin是一种酸性蛋白质，pI值为3.8，在pH 8.0-8.5和温度50摄氏度下发挥最佳作用。Religiosin的蛋白水解活性被PMSF和糜蛋白酶抑制剂强烈抑制，表明该酶是丝氨酸蛋白酶。Religiosin的消光系数（λ（1%）（280））为29.47 M（-1）cm（-1），每个分子含有16个色氨酸、26个酪氨酸和11个半胱氨酸残基。该酶显示出广泛的底物特异性对天然以及合成底物的表观K（m）为0.066 mM和6.25 mM，分别使用酪蛋白和Leu-pNA。MS/MS分析证实了该酶的新奇。对变性剂、金属离子和洗涤剂以及在宽pH和温度范围内，曲马红具有高度稳定性。此外，该酶还具有凝乳和洗涤剂活性。
The crude latex of Ficus religiosa is decolorized by activated charcoal. Decolorization follows the Freundlich and Langmuir equations. A serine protease, named religiosin, has been purified to homogeneity from the decolorized latex using anion exchange chromatography. Religiosin is a glycoprotein with a molecular mass of 43.4 kDa by MALDI-TOF. Religiosin is an acidic protein with a pI value of 3.8 and acts optimally at pH 8.0-8.5 and temperature 50 degrees C. The proteolytic activity of religiosin is strongly inhibited by PMSF and chymostatin indicating that the enzyme is a serine protease. The extinction coefficient (epsilon(1%)(280)) of religiosin is 29.47 M(-1) cm(-1)with 16 tryptophan, 26 tyrosine, and 11 cysteine residues per molecule. The enzyme shows broad substrate specificity against natural as well as synthetic substrates with an apparent K(m) of 0.066 mM and 6.25 mM using casein and Leu-pNA, respectively. MS/MS analysis confirms the novelty of the enzyme. Religiosin is highly stable against denaturants, metal ions, and detergents as well as over a wide range of pH and temperature. In addition, the enzyme exhibits milk-clotting as well as detergent activity.