以HPD-750大孔树脂为载体材料固定化果胶酶

Pectinase Immobilization Using HPD-750 Macroporous Resin as Carrier Material

HPD-750树脂是中极性大孔吸附树脂,生物相容性好,机械性能稳定,具有较大的比表面积,可用于固定化酶载体材料。本文以HPD-750大孔树脂为载体固定化果胶酶,研究各因素对固定化酶的影响,并采用正交试验对固定化条件进行优化。结果表明,当pH为4.0、固定化温度为45℃、固定化时间为4h、加酶量为0.16g/mL时,固定化酶活力可达5146U/mg。以HPD-750大孔树脂为载体材料制备的固定化酶相较于游离酶具有更好的酸碱稳定性和热稳定性。在循环使用10次后,酶活力依然保留80%以上;4℃储藏25d之后,其酶活力仍保留60%以上。与D311大孔树脂、聚丙烯酰胺和海藻酸钠微球制备的固定化酶相比,HPD-750大孔树脂固定化酶的活性、操作稳定性、机械稳定性和储存稳定性都较好。该结果说明,HPD-750大孔树脂可作为固定化酶较好的载体材料。

HPD-750 resin is a medium-polar macroporous adsorption resin with good biocompatibility, stable mechanical properties and large specific surface area, which can be used for immobilizing enzyme. In this paper, pectinase was immobilized by HPD-750 macroporous resin, and the effects of various factors on the immobilized enzyme were studied. The optimized immobilization parameters are: pH 4.0, the immobilization temperature 45 ℃, the immobilization time 4 h, and the amount of enzyme added 0.16 g/mL, Under these conditions, the immobilized enzyme activity can reach 5146 U/mg. Compared with free pectinase, the immobilized enzyme was found to exhibit better stability of pH and thermo. Additionally, the immobilized pectinase had remarkable reusability and storage stability, the enzyme activity remained more than 80% after 10 cycles of use;and after storage at 4 ℃ for 25 days, the enzyme activity remained more than 60%. Compared with immobilized enzyme prepared D311 macroporous resin, polyacrylamide and sodium alginate microspheres, the HPD-750 immobilized enzyme has better activity, operational stability, mechanical stability and storage stability. Hence that the HPD-750 macroporous resin could be used as a good carrier material for immobilized enzymes.