交联海藻糖合酶聚集体的制备及其性质

doi:10.7506/spkx1002-6630-201309024

食品科学 ›› 2013, Vol. 34 ›› Issue (9): 108-113.doi: 10.7506/spkx1002-6630-201309024

交联海藻糖合酶聚集体的制备及其性质

陈颖¹,肖辰鹏²,陈晓云¹,杨丽维¹,唐柳¹,李明春²,张峻¹

1.天津市林业果树研究所，天津 300384；2.南开大学生命科学学院，天津 300071

收稿日期:2012-11-06 修回日期:2013-03-15 出版日期:2013-05-15 发布日期:2013-05-07
通讯作者: 张峻 E-mail:zhangjun85wsw@yahoo.com.cn
基金资助:
国家自然科学基金面上基金项目(21076162)；天津市应用基础及前沿技术研究计划项目(10JCYBJC09600；10JCYBJC05000)

Preparation and Properties of Cross-linked Enzyme Aggregates of Trehalose Synthase

CHEN Ying1，XIAO Chen-peng2，CHEN Xiao-yun1，YANG Li-wei1，TANG Liu1，LI Ming-chun2，ZHANG Jun1,*

1. Tianjin Institute of Forestry and Pomology, Tianjin 300384, China；
2. College of Life Science, Nankai University, Tianjin 300071, China

Received:2012-11-06 Revised:2013-03-15 Online:2013-05-15 Published:2013-05-07
Contact: ZHANG Jun E-mail:zhangjun85wsw@yahoo.com.cn

摘要/Abstract

摘要：

采用交联酶聚集体技术制备了海藻糖合酶交联酶聚集体（CLEAs），研究了不同沉淀剂、酶与交联剂比例、交联强度及硼氢化钠还原处理对海藻糖合酶CLEAs活性的影响及其结构特征与反应性能。结果表明：采用10mg/mL的酶浓度以30%的PEG为沉淀剂，以终浓度0.5%的戊二醛室温交联2h，再以硼氢化钠还原处理后，制备得到的海藻糖合酶CLEAs最适催化温度为70℃，比游离酶提高20℃，最适pH为7.0，与游离酶相比，CLEAs的温度及pH稳定性均得到提高，对Zn2+、Cu2+、Fe2+、Al3+金属离子的抗性明显增强。微观形貌分析表明单个海藻糖合酶CLEA粒径在0.2-0.5μm，众多CLEA以“脚手架”结构形成大小不一的集簇。

关键词: 海藻糖合酶, 交联酶聚集体(CLEAs), 无载体固定化, 集簇

Abstract:

This paper describes the preparation and properties of cross-linked enzyme aggregates (CLEAs) of trehalose
synthase. The effects of different precipitants, enzyme-to-precipitant concentration ratio, cross-linking intensity and NaBH4
treatment on the activity of CLEAs were investigated as well as their structural features and reactivity. The optimum conditions
for CLEAs preparation were achieved when 10 mg/mL trehalose synthase solution was added with 30 mg/mL of PEG, crosslinked
with 0.5% glutaraldehyde at ambient temperature for 2 h, followed by NaBH4 reduction. CLEAs revealed optimum
reaction temperature of 70 ℃, 20 ℃ higher than that of the free enzyme, and optimum pH of 7.0. Compared to the free enzyme,
CLEAs indicated improved heat and pH stability. Moreover, the tolerance to some metal ions such as Zn2+, Cu2+, Fe2+ and Al3+
was remarkably enhanced. The microscopic morphology of CLEAs showed individual aggregates of 0.2—0.5 μm in particle
size and numerous clusters of different sizes in a “scaffolding” structure.

Key words: trehalose synthase (Tres), cross-linked enzyme aggregates (CLEAs), carrier-free immobilization, cluster

中图分类号:

Q814.2

陈颖肖辰鹏陈晓云杨丽维唐柳李明春张峻. 交联海藻糖合酶聚集体的制备及其性质[J]. 食品科学, 2013, 34(9): 108-113.

CHEN Ying1，XIAO Chen-peng2，CHEN Xiao-yun1，YANG Li-wei1，TANG Liu1，LI Ming-chun2，ZHANG Jun1,*. Preparation and Properties of Cross-linked Enzyme Aggregates of Trehalose Synthase[J]. FOOD SCIENCE, 2013, 34(9): 108-113.

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交联海藻糖合酶聚集体的制备及其性质

Preparation and Properties of Cross-linked Enzyme Aggregates of Trehalose Synthase

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