铜金属螯合载体固定糖化酶

摘要

[目的]制备出含Cu2+的琼脂糖-IDA螯合载体及对其固定糖化酶工艺条件进行优化.[方法]利用金属螯合配体(IDA-Cu2+)与蛋白质表面供电子氨基酸相互作用的原理制备载体,采用紫外分光光度法测定不同影响因素下固定化糖化酶的酶活.[结果]Cu2+的加入量和固定化过程的酸度比给酶量对固定化糖化酶的活性影响还要大,在给酶量80 mg/g载体、1.0× 10-2 mol Cu2+/g载体、pH 4.6和固定化4h的固定化条件下,固定化酶活为252.1 U/g,重复使用5次后酶活为首次固定化酶活的65.1％.[结论]该Cu2+-IDA-金属螯合琼脂糖可用于淀粉水解糖化酶的优良固定化载体材料.%[Objective] A chelate carrier with agarose containing Cu2+ iminodiacetic acid (IDA) for hydrolysis saccharification enzyme was prepared and the fixed the glucoamylase process conditions with the chelate carrier was optimized. [Methods] Based on the technology of protein separation with immobilized mental ion affinity chromatography, the method for immobilization of glucoamylase has been selected, and the enzyme activity of the immobilized glucoamylase was determined using the UV spectrophotometric for research the process influencing factors. [Results] The optimum immobilization conditions of enzyme were as flows: enzyme load of 80 mg/g carrier, Cu2+ of l.0×10-2 mol/g carriers, immobilized time of 4 h, pH value of 4.6, and the activity of immoblized glucoamylase was 252.1 U/g. The activity of im-moblized glucoamylase on the regenerated matrix keep 65.1% effective of the new enzyme activity after used 5 times. [Conclusion] The Cu2+-IDA-agarose magnetic metal chelate can be used excellent immobilization carriers for starch hydrolysis saccharification enzyme.