磷脂酶D在纤维素载体上的共价固定化研究.pdfVIP

磷脂酶D 在纤维素载体上的共价固定化研究 ? 赵彭花 张小里 赵彬侠 鱼园 何娜 (西北大学化工学院, 陕西 西安 710069) 摘要：以纤维素为载体、环氧氯丙烷（ECH ）为活化剂，共价固定化链霉菌发酵产磷脂酶D （PLD ）。考察 了载体活化及酶固定化的适宜条件；并研究了固定化酶的性能。结果表明，适宜的载体活化条件为：NaOH 1mol/L 20ml, 纤维素1g, ECH 4ml，室温，中速搅拌；适宜的酶固定化条件为：环氧基纤维素 1g，酶 30 ml （粗酶蛋白质浓度约0.8g/L ）、37℃、固定化反应时间 12h。获得的最大酶固定化率为48.8%，其固定化磷 脂酶的水解活力可达 10.5IU/g。固定化磷脂酶D 的最适pH 为 8.0、最适温度为50℃，与游离酶相比最适 pH 未改变，而最适温度提高10℃。固定化酶的热稳定性和操作稳定性均高于游离酶。 关键词: 磷脂酶D ；纤维素；共价固定化；环氧氯丙烷 中图分类号：Q81;Q814.2 文献标识码：A 文章编号： Covalence Immobilization of Phospholipase D on Cellulose ? ZHAO Penghua ，ZHANG Xiaoli ，ZHAO Binxia ，YU Yuan ，HE Na (College of Chemical Engineering， Northwest University, Xi’an 710069, Shaanxi, China) Abstract ：PhospholipaseD (EC3.1.4.4) from strepomyces was immobilized on carriers of cellulose by covalent coupling with epichlorohydrin(ECH) as intermediary linker. The optimization conditions for activation of cellulose and immobilition of enzyme was investigated, meanwhile, the character of immobilized PLD was studied. The results showed that the suitable conditions for the activation of the carrier are: NaOH 1M 20ml; Microcrystalline cellulose 1g; ECH 4ml; At room temperature; stir in the medium speed and the appropriate conditions for immobilize enzyme are: Epoxy cellulose 1g; PLD solution 30ml(crude protein 0.8g/L); 37℃; Immobilized reaction time 12h. The degree of immobilization can access to 48.8%. Compared with the free PLD, immobilized enzyme’s pH didn’t change while the temperature higher than free one by 10℃. The thermal and operational stability of immobilized PLD ar