微波辐射高效共价固定青霉素酰化酶

为提高青霉素酰化酶的共价固定化效率, 在微波辐射条件下将酶蛋白共价固定于介孔泡沫硅(MCFs)的孔道中. 通过正硅酸四乙酯水解缩合制备介孔泡沫硅, 再于微波辅助下将青霉素酰化酶共价固定在其孔道中. 以固定化酶相对活力和活力回收为指标, 考察了加酶量、固定化温度、微波辐射时间等条件对酶固定化效率的影响. 实验结果表明: 当加酶量为60 mg/g, 固定化温度为20 ℃, 微波辐射140 s, 固定化酶相对活力达到178.1%, 表观活力为1191.3 U/g(以湿重计). 与常规方法相比, 微波辅助固定化酶时, 固定化酶相对活力提高34.5%, 固定化时间亦大幅缩短至数分钟, 这为青霉素酰化酶的高效共价固定化提供了一条新的途径.

High Efficient and Covalent Immobilization of Penicillin Acylase under Microwave Irradiation

To enhance the penicillin acylase covalent immobilization efficiency, microwave irradiation was introduced to the enzyme immobilization process. Mesocellular siliceous foams (mesostructured cellular foams, MCFs) with large pores were synthesized by hydrolytic condensation of tetraethoxysilicone, and penicillin acylase was rapidly coupled to MCFs with covalent linkages by benzoquinine under microwave irradiation. The relative activity and the activity recovery of immobilized enzyme were taken as indexes, and the effects of immobilization conditions such as the reaction time, temperature and amount of penicillin acylase were investigated. When the amount of penicillin acylase, temperature and reaction time for immobilization was 60 mg/g MCFs, 20 ℃ and 140 s respectively, the activity of the wet immobilized enzyme was 1191.3 U/g, and the relative activity was up to 178.1%. Compared with those of conventional methods, the relative activity has increased by 34.5%, and the immobilization under microwave irradiation could shorten the reaction process to a few minutes, which provided a new way for immobilizing penicillin acylase.