摘要
以陶瓷为第一载体,壳聚糖为二次载体,戊二醛为交联剂,采用共价结合和吸附联用法制备固定化漆酶。考察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。结果表明,漆酶固定化的适宜条件为0.15g壳聚糖-陶瓷复合载体,加入3mL1.25mg/mL漆酶磷酸盐缓冲溶液(0.1mol/L,pH4.0),在4℃固定24h。酶的固定化效率是51.0%,固定化酶的酶活是55.87U/g,最适pH为3.0,最适温度分别为25℃和50℃。该固定化酶具有良好的贮存和操作稳定性。在pH3.0,温度25℃时,固定化酶对2,2-连氮基-双-(3-乙基苯并二氢噻唑啉-6-磺酸)二铵盐的表观米氏常数为66.64μmol/L。
Laccase has been immobilized on a ceramic-chitosan composite support by using glutaraldehyde as cross-linking reagent. The effects of varying the immobilization conditions were investigated and the properties of the immobilized enzyme were characterized. The optimal immobilization conditions were found to be： 3.0 mL of 1.25 mg/mL of laccase in phosphate buffer （0.1 mol/L, pH 4.0） reacted with 0.15 g ceramic-chitosan composite support at 4℃ for 24 h. Under these immobilization conditions, an immobilization efficiency of 51.0% was obtained. The maximum activity of the immobilized enzyme was 55.87 U/g. The immobilized enzyme exhibited its maximum activity at pH 3.0 and the activities of the immobilized enzyme were highest at 25℃ and 50 ℃. The Km value of immobilized laccase with diammonium 2, 2-azino-bis（3-ethylbenzothiazoline-6-sulfonate） was 66.64/-tmol/L in pH 3.0 buffer at 25℃. The thermal, operational, and storage stabilities of free laccase were all increased after immobilization.
出处
《北京化工大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第4期84-88,共5页
Journal of Beijing University of Chemical Technology(Natural Science Edition)
基金
燕山石化研究院技术开发项目(H2004195)
关键词
陶瓷-壳聚糖
固定化
漆酶
酶活力
ceramic-chitosan composite support
immobilization
laccase
enzyme activity
