酪氨酸酶的固定化及其对水中酚类物质的去除详解.docxVIP

 PAGE \* MERGEFORMAT 31 酪氨酸酶的固定化及其对水中酚类物质的去除 摘要 本文制备了不同质量比的壳聚糖/有机累托石插层复合物（CTS/OREC），并以CTS/OREC为载体通过吸附法和交联法固定酪氨酸酶。结果表明，吸附法固定化酶（A-TYR）的最佳条件为以壳聚糖/有机累托石质量比4:1的插层复合物为载体，pH值5.0，酶与载体质量比30 mg/g，固定化时间6h，制备得固定化酪氨酸酶的载酶量为17.30mg/g，单位载体酶活为11.67kU/g，酶活回收率87.9%；戊二醛交联法固定化酶（C-TYR）的最佳条件为以壳聚糖/有机累托石质量比为4:1的插层复合物为载体，pH值4.0，酶与载体质量比30 mg/g，固定化时间4 h，制备得固定化酪氨酸酶的载酶量为26.52mg/g，单位载体酶活为6.14kU/g，酶活回收率29.7%。固定化酶比游离酶有更好的操作稳定性和储存稳定性。4℃储存20天后A-TYR相对酶活为63.9%，C-TYR相对酶活为89.6%。A-TYR去除苯酚的最佳反应条件为pH 7.0、温度40℃；A-TYR去除4-氯酚的最佳反应条件为pH 5.0、温度30℃；A-TYR去除2,4-二氯酚的最佳反应条件为pH 6.0、温度30℃。A-TYR在最佳条件下完全去除苯酚、4-氯酚和2,4-二氯酚时的最佳酶与底物质量比分别为3.50mg/mg、1.38mg/mg、1.38mg/mg。 关键词：酪氨酸酶；固定化；壳聚糖；累托石；插层复合物；酚 Immobilization of tyrosinase and removal of phenolic compounds in water Abstract In this paper, different mass ratio of chitosan (CTS) and organic rectorite (OREC) was used to prepared intercalation composite (CTS/OREC) and CTS/OREC with mass ratio of 4:1 was used as a carrier to immobilize tyrosinase through adsorption and cross-linking method. The optimal immobilization condition of tyrosinase for adsorption (A-TYR) was pH 5, the mass ratio of enzyme?to?support 30 mg/g, immobilization time 6 h. In this condition, the protein loading yield of A-TYR was 17.30mg/g, enzyme activity of the unit support 11.67kU/g, and activity recovery 87.9%. The optimal immobilization condition of tyrosinase for glutaraldehyde cross-linking (C-TYR) was pH 4, the mass ratio of enzyme?to support 30 mg/g, immobilization time 4 h. In this condition, the protein loading yield of C-TYR was 26.52mg/g, enzyme activity of the unit support 6.14kU/g, and activity recovery 29.7%. The immobilized tyrosinase retained higher relative activity when the temperature and pH value changed, compared with the free enzyme. The immobilized tyrosinase have batter storage stability than the free enzyme, the A-TYR and C-TYR still retained 63.9% and 89.6% relative activity, respectively, even after 20 days stored in 4℃. A-TYR was used to remove phenol, 4-chlorophenol and 2,4–