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

作　　者：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] 

机构地区：[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

出　　处：《食品科学》2013年第9期108-113,共6页Food Science

基　　金：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)

摘　　要：采用交联酶聚集体(CLEAs)技术制备交联海藻糖合酶聚集体,研究不同沉淀剂、酶与交联剂浓度比例、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:采用质量浓度为10mg/mL的酶以质量浓度为30mg/mL的聚乙二醇(PEG)为沉淀剂,以体积分数为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值为7.0,与游离酶相比,CLEAs的温度及pH值稳定性均得到提高,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聚集体粒径在0.2～0.5μm,众多聚集体以"脚手架"结构形成大小不一的集簇。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.

关 键 词：海藻糖合酶 交联酶聚集体(CLEAs) 无载体固定化 集簇 

分 类 号：TS201.3[轻工技术与工程—食品科学]