凝胶法固定乙酰胆碱酯酶的研究

Study on Gel-immobilization Method of Acetylcholinesterase

以海藻酸钠为载体包埋固定乙酰胆碱酯酶(AChE),考察了影响酶固定化的重要因素,获得了最佳固定化条件。实验结果表明,将1U乙酰胆碱酯酶,2μLφ(戊二醛)为5%的戊二醛,1μL10g/L的牛血清白蛋白(BSA),与30g/L的海藻酸钠配成300μL的酶溶液,滴入20g/L的氯化钙溶液中,3℃下固定8h,制备的凝胶酶珠机械强度高,活力重现性好。在20g/L的氯化钙溶液中3℃下保存40d,活力基本不变,相对标准偏差为5.53%～6.82%。

Using Sodium alginate as a carrier, a immobilization method of acetylcholinesterase is reported. The important factors affecting immobilization are explored, and the optimum conditions are obtained. The emperimental results show that 300 μL enzyme solution, is composed of 1 U of acetylcholinesterase, 2 μL 5%(v/v)of glutaraldehyde, 1 μL 10 g/L of bovine serum albumin and 30 g/L of sodium alginate, which is dropped into 20 g/L of calcium chloride, and is saved for 8 h at 3 ℃.Using the described method, the enzyme grain can be produced. The activity of enzyme grain is quite similar. It may be saved in 20 g/L of calcium chloride for 40 d (RSD is 5.53%～6.82%).