Department of Applied Chemistry, Zhejiang Gongshang University, No. 149 Jiaogong Road, Hangzhou 310012, People's Republic of China.
Department of Applied Chemistry, Zhejiang Gongshang University, No. 149 Jiaogong Road, Hangzhou 310012, People's Republic of China.
Int J Biol Macromol. 2018 Jun;112:1175-1182. doi: 10.1016/j.ijbiomac.2018.02.079. Epub 2018 Feb 15.
A new type of magnetic metal chelating carrier (PCMM-IDA-Cu) was prepared for the immobilization of papain, using chitosan as raw material, nano FeO as magnetic material, SiO as porogen, iminodiacetic acid (IDA) as a chelating ligand, and binding with transition metal ion (Cu). The resulting products were well characterized by optical microscopy, scanning electron microscopy (SEM), vibrating sample magnetometer (VSM), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). The BBD (Box-Behnken Design) of RSM (Response Surface Methodology) was applied to analyze the optimum enzyme immobilization conditions. The results showed that the enzyme immobilization capacity was 94.18mg/g of PCMM-IDA-Cu, with 7.976U/mg of relative immobilized enzyme activity under the optimum conditions (pH6.73, 1.56mg enzyme/15.0mg carrier, 30.9°C), and the recovery of enzyme activity was reached 87.21%. Compared with the free papain, the immobilized papain displayed enhanced enzyme activity, superior enzymatic properties, good operational stability and reusability. It is worth mentioning that the novel carriers exhibited selectively biological adsorption capacity, and this technique is an alternative method for the immobilization of enzyme, making the process more efficient and economic.
一种新型的磁性金属螯合载体（PCMM-IDA-Cu）被制备用于固定木瓜蛋白酶，以壳聚糖为原料，纳米 FeO 为磁性材料，SiO 为致孔剂，亚氨基二乙酸（IDA）为螯合配体，并与过渡金属离子（Cu）结合。通过光学显微镜、扫描电子显微镜（SEM）、振动样品磁强计（VSM）、傅里叶变换红外光谱（FTIR）和热重分析（TGA）对所得产物进行了很好的表征。RSM（响应面法）的 BBD（Box-Behnken Design）被应用于分析最佳酶固定化条件。结果表明，在最佳条件下（pH6.73、1.56mg 酶/15.0mg 载体、30.9°C），酶固定化容量为 94.18mg/g 的 PCMM-IDA-Cu，相对固定化酶活性为 7.976U/mg，酶活回收率达到 87.21%。与游离木瓜蛋白酶相比，固定化木瓜蛋白酶显示出增强的酶活性、优越的酶学性质、良好的操作稳定性和可重复使用性。值得一提的是，新型载体表现出选择性的生物吸附能力，该技术是酶固定化的替代方法，使过程更高效、更经济。
