磁性聚乙烯醇微球固定化α-乙酰乳酸脱羧酶  

α-Acetolactatedecarboxylase immobilized onto magnetic polyvinyl alcohol microspheres

作　　者：钱斯日古楞 王红英[1] 刘阳[1] 耿金峰[1] 朱婧莹[1] 崔茂欣[1] 

机构地区：[1]大连工业大学生物与食品工程学院,辽宁大连116034

出　　处：《化工学报》2007年第8期2062-2066,共5页CIESC Journal

基　　金：大连市科技攻关项目(2004B3SF175)~~

摘　　要：Magnetic polyvinyl alcohol (PVA) microspheres with 8—64 μm in diameter were prepared from PVA by dispersion and copolymerization with Fe3O4 as magnetite.There were more functional groups, such as hydroxyl and carboxyl etc. on the microsphere surface.1,1′-Carbonyldiimidazole (CDI), a carbonylating agent, was used for the activation of hydroxyl groups of PVA, and α-acetolactatedecarboxylase (ALDC) was immobilized onto the magnetic PVA microspheres by covalent bonding through the amino group.The results showed that total activity, protein binding, specific activity and activity retention of the immobilized enzyme were 63293 U·g -1, 72.67 mg·g -1, 870.96 U·mg -1 and 48.71%,respectively.The optimum temperature of immobilization enzyme was 50℃ and the optimum pH was 6.0.Compared with the free enzyme of ALDC, the thermal, operational and pH stability of the immobilized enzyme were improved.After being stored at 4℃, pH 6.0 for 31 days, the immobilized ALDC retained 95.7% of its initial activity which was 8% higher than the free enzyme.Magnetic polyvinyl alcohol （PVA） microspheres with 8--64 p.m in diameter were prepared from PVA by dispersion and copolymerization with Fe304 as magnetite. There were more functional groups, such as hydroxyl and carboxyl etc. on the microsphere surface. 1, 1＇-Carbonyldiimidazole （CDI）, a carbonylating agent, was used for the activation of hydroxyl groups of PVA, and α- acetolactatedecarboxylase （ALDC） was immobilized onto the magnetic PVA microspheres by covalent bonding through the amino group. The results showed that total activity, protein binding, specific activity and activity retention of the immobilized enzyme were 63293 U·g^-1, 72.67 mg·g^-1, 870. 96 U· mg^-1 and 48.71%, respectively. The optimum temperature of immobilization enzyme was 50℃ and the optimum pH was 6.0. Compared with the free enzyme of ALDC, the thermal, operational and pH stability of the immobilized enzyme were improved. After being stored at 4℃, pH 6.0 for 31 days, the immobilized ALDC retained 95.7% of its initial activity which was 8% higher than the free enzyme.

关 键 词：磁性微球 聚乙烯醇 Α-乙酰乳酸脱羧酶 固定化酶 

分 类 号：Q814.2[生物学—生物工程]