D301树脂固定化假丝酵母脂肪酶

Immobilization of Candida sp. lipase on resin D301

摘要:

本研究选择7种吸附和离子交换树脂进行了假丝酵母脂肪酶(Candida sp. lipase)的固定化试验, 通过测定固定化后各脂肪酶的酶活, 筛选出固定化效果较好的弱碱性阴离子交换树脂D301; 并通过扫描电镜将D301与脂肪酶Novozym 435的表面形貌做比较, 进一步选定D301树脂作为载体, 并对其采用戊二醛交联固定化, 研究并优化了其固定化条件。结果表明, 5%戊二醛溶液的加入量为8 mL, 处理时间为5 h, 酶液浓度为1.0 g/L, 磷酸缓冲盐溶液pH 6.0, 固定化处理10 h效果最好, 获得的固定化酶活力可达35 U/mg, 酶的固定化效率约为3.5 U/(mg·h)。

关键词:D301, 树脂, 固定化, 假丝酵母脂肪酶

Abstract:

We immobilized Candida sp. lipase onto seven kinds of industrial adsorption and ion exchange resins. By determining the activity of each immobilized enzyme, the weakly basic anionic exchange resin of D301 showed the best results for the immobilization of Candida sp. lipase. Comparing the scanning electron micrographs of D301 with Novozym 435 (immobilized Candida antarctica lipase B from Novo Nordisk Corp.), we selected D301 as a carrier for the immobilization of Candida sp. lipase. And we pretreated the resin D301 with the bifunctional agent glutaraldehyde and crosslinked it with Candida sp. lipase. The optimal conditions for the immobilization of Candida sp. lipase were as follows: 8 mL of the amount of 5% glutaraldehyde solution, five hours of the time pretreated D301 with glutaraldehyde, 1.0 g/L the concentration of Candida sp. lipase used, pH of the phosphate buffered, 6.0 and 10 hours of time for immobilization, respectively. The activity of immobilized enzyme was over 35 U/mg and the efficiency of immobilization was around 3.5 U/(mg·h).

Key words:D301, resins, immobilization, Candida sp. lipase