为寻求一种廉价高效的漆酶固定化方法,以树脂D380为载体、戊二醛为交联剂,结合吸附法和交联法固定真菌漆酶,并研究固定化漆酶的性质。最优固定化条件:1 g载体在20 mL 0.7%的戊二醛溶液中交联2 h后,加入2 mL稀释酶液和5 mL pH为4的醋酸-醋酸钠缓冲溶液,在25℃下吸附反应6 h,最终获得的酶活回收率为65.94%。固定化漆酶的最适温度为20℃,最适pH为4,在4℃下保存14 d仍保持约80%的酶活力。自由漆酶和固定化漆酶的Km值分别为1.207mmol/L和1.616 mmol/L,说明固定后漆酶对底物的亲和力有所下降。
【英文摘要】
In order to seek a cheap and efficient way for immobilization of laccase,resin D380 as support and glutaraldehyde(GLU) as cross-linking agent were employed for immobilization of fungal laccase combining adsorption and cross-linking methods,and the properties of the immobilized laccase were also studied.The optimum conditions of the immobilization are as follows: after cross-linking for 2 h with 20 mL GLU of 0.7%,1 g support were conducted in 2 mL diluent laccase solution and 5 mL HAC-NaAC buffer solution(pH...
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