交联草酸脱羧酶聚集体的制备及其性质

食品科学 ›› 2013, Vol. 34 ›› Issue (1): 215-219.

交联草酸脱羧酶聚集体的制备及其性质

梁跃1，林日辉1,*，黄文勤2，秦梦1，李香香1，何美卿1，郑元博3

1.广西民族大学化学化工学院，化学与生物转化过程新技术广西高校重点实验室 2.南宁奕德环境科技有限公司 3.东北大学理学院

收稿日期:2011-11-11 修回日期:2012-12-07 出版日期:2013-01-15 发布日期:2013-01-07
通讯作者: 林日辉 E-mail:linrihui_0@yahoo.com.cn
基金资助:
广西化工过程创新技术研发平台建设，国家民委科研项目资助课题。

Preparation and Properties of Cross-Linked Oxalate Decarboxylase Aggregates

Received:2011-11-11 Revised:2012-12-07 Online:2013-01-15 Published:2013-01-07

摘要/Abstract

摘要： 为了获得缓解泌尿系统草酸盐结石病症的酶制剂，用无载体固定化方法——交联酶聚集体(CLEAs)制备交联草酸脱羧酶聚集体。使用基因工程菌E.coli BL21(DE3)/pET32a/YvrK诱导表达制备草酸脱羧酶粗酶液，用30%的乙醇进行沉淀分离提纯，纯化倍数为2.7倍，酶活回收率91.2%；在戊二醛添加量为0.06%、pH5、牛血清白蛋白添加量0.5g/L、4℃的条件下处理该酶2h，得到交联草酸脱羧酶聚集体(oxdc-CLEAs)，酶活回收率达95.4%；酶学性质研究表明，相对游离草酸脱羧酶，交联草酸脱羧酶聚集体的耐酸性、耐热性、耐胰蛋白酶降解能力均有提高。

关键词: 草酸脱羧酶, 交联酶聚集体(CLEAs), 制备, 性质

Abstract: Cross-linked oxalate decarboxylase aggregates (oxdc-CLEAs) for use as an enzyme preparation to relieve urinary oxalate stone disease were prepared by a carrier-free immobilization method. Crude enzyme solution was obtained from the induced expression of the genetically engineered strain E.coli BL21(DE3)/pET32a/YvrK, and oxalate decarboxylase was separated by adding 30% ethanol with a purification factor of 2.7 and an activity recovery of 91.2%. Cross-linked oxdc-CLEAs were further obtained after 2 h of cold treatment at 4 ℃ in the presence of 0.5 g/L bovine serum albumin (BSA) and 0.06% glutaraldehyde at pH 5, resulting in an activity recovery of 95.4%. Enzymatic characterization showed that cross-linked oxdc-CLEAs had improved tolerance to acid, heat and trypsin degradation when compared to free oxalate decarboxylase.

Key words: oxalate decarboxylase, cross-linked enzyme aggregates, preparation, properties

中图分类号:

Q814.4

梁?跃1，林日辉1,*，黄文勤2，秦?梦1，李香香1，何美卿1，郑元博3. 交联草酸脱羧酶聚集体的制备及其性质[J]. 食品科学, 2013, 34(1): 215-219.

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