交联海藻酸钠固定化柚(皮)苷酶
Immibilization of naringinase onto cross-linked alginate sodium carrier
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摘要: 以海藻酸钠为载体,戊二醛为交联剂,采用交联-包埋-交联法对柚(皮)苷酶进行了固定化。在单因素实验基础上,通过正交实验得到海藻酸钠固定化柚(皮)苷酶的最优工艺条件:海藻酸钠质量浓度3.0%,给酶量为0.01mg/g载体,前交联戊二醛体积分数2.0%,前交联时间1.5h,后交联戊二醛体积分数0.025%,后交联时间2h,制备的固定化酶最高活力5.07U/g。同时,对固定化柚(皮)苷酶的稳定性进行了研究,结果表明:固定化酶的温度耐受性与存储稳定性较游离酶有较大幅度的提高;固定化酶重复使用7次(60℃,pH4.0)后,活力仍然保持在60%。Abstract: Naringinase was immobilized into alginate sodium matrices with glutaraldehyde via crosslinked- entrapment- crosslinked technique. According to the orthogonal test design based on the single factor experiments, the optimal immobilization conditions were obtained as follows: the mass concentration of alginate sodium 3.0%, the amount of enzyme onto supports 0.01mg /g, the previous crosslinking volume fraction of glutaraldehyde 2.0%, the previous crosslinking time 1.5h, the after crosslinking volume fraction of glutaraldehyde 0.025%, the after crosslinking time 2h, and the highest activity of immobilized naringinase was 5.07U /g.Meanwhile, operational properties of the immobilized naringinase were investigated. The results showed that the temperature tolerance and storage stability of immobilized naringinase had dramatically increased than the free enzyme. The immobilized naringinase retained about 60% of initial activities after 7 recycles ( 60℃, pH4.0) .