摘要
以磁性壳聚糖微球为载体,戊二醛为交联剂,共价结合制备固定化漆酶。探讨了漆酶固定化的影响因素,并对固定化漆酶的性质进行了研究。确定漆酶固定化适宜条件为50mg磁性壳聚糖微球,加入10mL0.8mgmL漆酶磷酸盐缓冲液(0.1molL,pH7.0),在4℃固定2h。固定化酶最适pH为3.0,最适温度分别为10℃和55℃,均比游离酶降低5℃。在pH3.0,温度37℃时,固定化酶对ABTS的表观米氏常数为171.1μmolL。与游离酶相比,该固定化漆酶热稳定性明显提高,并具有良好的操作和存储稳定性。
Laccase was immobilized on magnetic chitosan microspheres by using glutaraldehyde as cross-linking reagent. The immobilization conditions and characterization of the immobilized enzyme were investigated. The optimal immobilization conditions for laccase were: 10mL of 0.8mg/mL of laccase in phosphate buffer(0.1mol/L,pH 7.0) reacted with 50mg of magnetic chitosan microspheres at 25℃ for 1h and subsequently was kept at 4℃ for 2h. The immobilized enzyme exhibited the maximal activity at pH 3.0.The optimal temperatures for immobilized enzyme were 10℃ and 55℃. The K-m value of immobilized laccase for ABTS was 171.1μmol/L in pH 3.0 phosphate buffer at 37℃. Compared with free enzyme ,the thermal, operational, and storage stabilities of the enzyme were increased after the immobilization.
出处
《微生物学报》
CAS
CSCD
北大核心
2005年第4期630-633,共4页
Acta Microbiologica Sinica
基金
国家自然科学基金资助项目(60377032)~~
关键词
磁性壳聚糖微球
漆酶
固定化
稳定性
Chitosan, Immobilization, Laccase, Immobilization, Stability
