烟草多酚氧化酶的分离与固定化技术研究

多酚氧化酶属于氧化还原酶类，国际酶学委员会推荐名为儿茶酚氧化酶（ＥＣ１．１０．３．１ｐｏｌｙｐｈｅｎｏｌｏｘｉｄａｓｅ，ＰＰＯ）．该酶与食品工业、三废处理、医药卫生关系较为密切，因而研究较多．如近年来鸭梨［１］、蘑菇［２］、香蕉果肉组织［３］、荔枝果皮［４］等等中的多酚氧化酶均有研究报道．目前研究用固定化多酚氧化酶检测废水中酚类物质含量，进行环境检测；及其从工业废水中除去酚类，达到治理三废的目的．Ｍｏｓｂａｃｎ［５］（１９７６）研制成多酚氧化酶固定化酶柱，与氧电极检测器组合联用，可检测水中２０…

Study on Isolation and Immobilization of Polyphenol Oxidase

Polyphenol oxidase was purified from leaves of Nicotiana tobaccum and the needle shape crystals were formed. The enzyme was immobilized on nylon membrane with glutaraldehyde as the cross linking agent. The appropriate conditions for immobilization were as follows: (1) The concentration of enzyme, 13 5 U/ml (concentration of protein: 0 03 mg/ml); (2) The optimum time for cross linking by glutaraldehyde was 20 minutes; (3)The optimum time for immobilization was 7 hours; (4) The optimum concentration of glutaraldehyde was 0 25%; (5) The optimum temperature was 4℃. The activity of immobilized enzyme reached 35—40 U/g matrix, and the activity recovery was 76 2% and the coupling efficiency was 76 3%. The stability of immobilized enzyme under acid, basic and high temperature conditions were enhanced and shifted toward basic pH range. It will be beneficial for industrial utilization.