改良壳聚糖固定化真菌漆酶的研究

Studies on Fungal Laccase Immobilized by Improved Chitosan

以改良壳聚糖-壳聚糖铜为载体,戊二醛为交联剂对真菌漆酶进行了固定化研究,探讨了其固定化条件及固定化酶的部分性质。结果表明:戊二醛最适浓度为0.5%,交联最佳时间14 h,最佳给酶量约1.3 mg,固定时间12 h或0.5 h。在此条件下获得的固定化漆酶酶活回收率分别为56.4%与51.2%。与游离酶相比,固定化漆酶与作用底物邻联甲苯胺的亲和力降低,但固定化漆酶的稳定性有明显改善。最适温度分别为40℃和80℃,分别比游离酶提高10℃和50℃;90℃条件下保温5 h后固定化酶活保留率分别为57.4%和40%,而相同条件下游离酶酶活明显下降。

Laccase from Amanita verng was immobilized on chitosan complexed by Cu^＋ by means of covalent coupling to a glutaradehyde-pretreated support. The condition for immobilizing and part of the qualities of the immobilized enzyme was studied. The optital conditions of immobilization could be specified as： 0.5%glutaradehyde crosslinked for 14 h, added about 1.3 mg free enzyme to immobilized for 12 h or 0.5 h, the recovery of which were 56.4% and 51.2%. Compared with the free enzyme, the affinity of immobilized enzymes decreased to otoluidine while the stability of them was greatly improved. The optital temperature were 40℃ and 80℃, semperately improving 10℃ and 50℃ ;placed in 90℃ for 5 h, the remaining activity of the immobilized enzyme were 57.4% and 40% ,while that of the free enzyme obviously decreased.