青霉素酰化酶在新型复合载体上的固定化研究

Studies on immobilization of penicillin acylase on novel complex carrier PAM/silica gel

在反应温度为45℃、反应时间为6 h、丙烯酰胺(AM)质量浓度为40 g/L、引发剂用量为单体质量1%的条件下制得PAM-SiO2复合载体,利用红外光谱表征了其化学结构,热失重法测定其接枝量为21%。在所制PAM-SiO2复合载体上固载青霉素酰化酶,其固载最适条件为:戊二醛用量0.1 mmol/L,pH值7.64,给酶量1%,温度42℃,时间11 h^12 h。此条件下所得固定化酶的活力为34000 U/mL;测得所制固定化酶的最适pH值为7.82,最适温度为45℃。

The PAM-SiO2 complex vector with 21 percent of the grafting yield was obtained under the optimal conditions of 45 ℃,reaction time about 6 h,AM concentration 40 g/L,initiator dosage of 1% of monomer.The chemical composition and structure of the composite carrier were characterized via IR.This vector was used to immobilize penicillin acy lase.The optimal immobilizing conditions is 1% glutaraldehyde,pH 7.64,enzyme concentration is 1%,temperature is 42 ℃ and time is 11 h～12 h.After that we get a immobilized enzyme which activity was 34 000 U/mL.Finally,the optimum conditions of the immobilized enzyme system are pH 7.82 and 45 ℃.