Chaitanyakumar Amballa, Anbalagan M
School of Bio-Sciences and Technology, VIT University, Vellore, 632014, India.
AMB Express. 2016 Dec;6(1):89. doi: 10.1186/s13568-016-0261-5. Epub 2016 Oct 4.
Enzymes find their applications in various industries, due to their error free conversion of substrate into product. Tannase is an enzyme used by various industries for degradation of tannin. Biochemical characterization of a specific enzyme from one organism to other is one of the ways to search for enzymes with better traits for industrial applications. Here, tannase encoding gene from Staphylococcus lugdunensis was cloned and suitability of the enzyme in various conditions was analysed to find its application in various industry. The recombinant protein was expressed with 6× His tag and purified using nickel affinity beads. The enzyme was purified up to homogeneity, with approximate molecular weight of 66 kDa. Purified tannase exhibited specific activity of about 716 U/mg. Optimum enzyme activity was found to be 40 °C at pH 7.0. Biochemical characterization revealed; metal ions such as Zn, Fe, Fe and Mn inhibited tannase activity, and SDS at lower concentration, increased tannase activity. Non polar organic solvents increased the tannase activity and polar solvents inhibited the tannase activity. Tannase immobilization studies show protection of the enzyme under wide range of pH and temperature. Also in this study we report a method for recovery and repeated use of the tannase.
由于酶能将底物无差错地转化为产物，因此在各种行业中都有应用。单宁酶是一种被各行业用于降解单宁的酶。对一种生物的特定酶与另一种生物的酶进行生化特性分析，是寻找具有更适合工业应用特性的酶的方法之一。在此，克隆了来自路邓葡萄球菌的单宁酶编码基因，并分析了该酶在各种条件下的适用性，以确定其在各行业中的应用。重组蛋白用6×His标签表达，并用镍亲和珠进行纯化。该酶被纯化至同质，近似分子量为66 kDa。纯化后的单宁酶表现出约716 U/mg的比活性。发现最适酶活性在pH 7.0、40℃条件下。生化特性分析表明；锌、铁、铁和锰等金属离子会抑制单宁酶活性，较低浓度的十二烷基硫酸钠会增加单宁酶活性。非极性有机溶剂会增加单宁酶活性，而极性溶剂会抑制单宁酶活性。单宁酶固定化研究表明该酶在较宽的pH和温度范围内受到保护。此外，在本研究中我们还报告了一种单宁酶回收和重复使用的方法。
