有机磷降解酶的固定化及其工业化应用初探

Immobilization of organophosphorus hydrolase and pilot study of its industrialisation application

采用一种新型的酶固定化方法，在大孔微球中对有机磷降解酶（OPHC2）固定化，制备固相载体化的交联酶聚集体固定化酶。对具有实际应用意义的参数：热稳定性、牢固度和连续反应批次进行了重点研究。结果表明，有机磷降解酶固定化酶的半衰期在70℃时比游离酶提高了2．1倍。固定化酶分别在37℃的摇床中经14h高速振荡，以及在pH8、37℃条件下循环反应49个批次降解甲基对硫磷后，均未发现固定化酶酶活力下降。在37℃，有机磷降解酶固定化酶柱以每小时3倍柱床体积的速率对甲基对硫磷农药乳油的稀释液（甲基对硫磷含量为216mg／L）进行降解处理，系统连续工作156h，甲基对硫磷的降解率保持在98．4％～99．9％。

Organophosphrous hydrolase （OPHC2） were immobilized in macroporous microsphere by the novel method for the enzyme immobilization, and prepared the immobilized enzyme which supported on solid state by cross-linking of enzyme aggregates （CLEAs）. The practical parameters： thermal stability, physical stability and the continuous reaction ablility were studied. The results showed that the half life of immolibilized enzyme was 2. 1 times more than native enzyme at 70 ℃. The activity of immolibized enzyme was not lessened after the immolibized enzymes had been shaked in shaker for 14 hours at 37 ℃ and hydrolyzed methyl parathion continuously for 49 batches at 37 ℃ （pH 8）. The hydrolytic rates of methyl parathion were 98.4% - 99. 9% in the immobilized enzymes column when it hydrolyzed the diluent solution （216 mg/L methion parathion） of 50% methyl parathion at the speed of 3 times of the column bed for 156 hours at 37 ℃.