功能基化聚丙烯酸甲脂固定化氨基酰化酶的研究

Studies on Immobilization of Aminoacylase on Functionalized Polvmethvl Acrvlate

以功能基化聚丙烯酸甲酯为载体制备固定化氨基酰化酶时,载体制备时的交联度,致孔剂用量和混合致孔剂比例均会影响酶的固定化效率。结果表明25%交联度和100%致孔剂用量制备的载体最佳。用此载体固定化氨基酰化酶可提高酶对高底物浓度、离子强度、温度、pH、有机溶剂和蛋白质变性剂的耐受能力。用固定化酶柱式反应器连续拆分N-乙酰基-DL-苯丙氨酸,使用一个月仍可保留90%的酶活力。

Functionalized polymethyl acrylate was used for immobilization of aminoacylase from Aspergilhs oryzae. Effects of degree of cross-linking, amount of porogenic agent and mixed porogenic agents on the activity of immobilized aminoacylase were investigated. The results showed the activity of immobilized aminoacylase was the highest when the carrier was 25% cross-linking and 100% porogenic agent.Effects of substrate concentration, ionic concentration, temperature and pH on immobilized aminoacylase were compared with those on soluble aminoacylase and effects of some organic solvents and protein denaturants on immobilized aminoacylase were investigated. A column reactor packed with immobilized aminoacylase was used to resolve continuously N-acyl-DL-phenylalanine for a month, and 90% activity of immobilized aminoacylase was retained.