木瓜蛋白酶交联聚体的制备及性质

高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (9): 1774.

木瓜蛋白酶交联聚体的制备及性质

王梦凡, 齐崴, 苏荣欣, 何志敏

天津大学化工学院化学工程研究所, 化学工程联合国家重点实验室, 天津 300072

收稿日期:2009-12-14 出版日期:2010-09-10 发布日期:2010-09-10
通讯作者: 齐崴, 女, 博士, 教授, 博士生导师, 主要从事酶工程领域研究. E-mail: qiwei@tju.edu.cn
基金资助:
国家自然科学基金(批准号: 20976125)、国家“八六三”计划项目(批准号: 2008AA10Z318)、教育部新世纪优秀人才(批准号: NCET-08-0386)、教育部科学技术研究重点项目(批准号: 108031)和天津市应用基础研究计划项目(批准号: 10JCYBJC05100)资助.

Preparation and Properties of Cross-linked Enzyme Aggregates of Papain

WANG Meng-Fan, QI Wei^*, SU Rong-Xin, HE Zhi-Min

State Key Laboratory of Chemical Engineering(Tianjin University), Chemical Engineering Research Center, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China

Received:2009-12-14 Online:2010-09-10 Published:2010-09-10
Contact: QI Wei. E-mail: qiwei@tju.edu.cn
Supported by:
国家自然科学基金(批准号: 20976125)、国家“八六三”计划项目(批准号: 2008AA10Z318)、教育部新世纪优秀人才(批准号: NCET-08-0386)、教育部科学技术研究重点项目(批准号: 108031)和天津市应用基础研究计划项目(批准号: 10JCYBJC05100)资助.

摘要/Abstract

摘要： 采用交联酶聚体(CLEAs)技术制得了木瓜蛋白酶CLEAs, 优化了制备条件. 以纯乙醇为蛋白沉淀剂, 质量分数40%戊二醛为交联剂, 于4 ℃下对酶沉淀聚体交联16 h; 所得木瓜蛋白酶CLEAs的最适pH为6.0(游离酶最适pH=7.0), 最适温度范围由游离酶的80 ℃拓宽为50～80 ℃, 热稳定性和溶液稳定性亦明显提高; 微观形貌分析证明木瓜蛋白酶CLEAs优良的催化效能及稳定性来自于CLEAs单元所具有的高比表面积及单元内部多点共价固定的结合方式.

关键词: 交联酶聚体, 固定化, 木瓜蛋白酶, 稳定性, 形貌, 集簇

Abstract: Cross-linked enzyme aggregates(CLEAs) is a non-carrier immobilization technique with the advantages of simple preparation, low cost and high unit activity. In this paper, papain-CLEAs were prepared and the effects of precipitation and cross-linking on CLEAs activity were investigated. It is shown that the highest activity of papain-CLEAs were obtained when using pure ethanol as precipitant, 40% glutaraldehyde as cross-linker and cross-linking at 4 ℃ for 16 h. Moreover, the optimal catalytic conditions and operational stabilities of papain-CLEAs were characterized. Papain-CLEAs shifted the optimal pH from 7.0 to 6.0 and broaden the optimal temperature to the range of 50—80 ℃. Papain-CLEAs also enhanced thermal and storage stabilities compared with free enzyme. It is indicated from the morphology that these improved performances resulted from the high surface area and multi-points covalent cross-linking structure of every single papain-CLEAs. These improved characters show practical advantages of papain-CLEAs in biocatalysis.

Key words: Cross-linked enzyme aggregate, Immobilization, Papain, Stability, Morphology, Cluster

TrendMD:

王梦凡, 齐崴, 苏荣欣, 何志敏. 木瓜蛋白酶交联聚体的制备及性质. 高等学校化学学报, 2010, 31(9): 1774.

WANG Meng-Fan, QI Wei*, SU Rong-Xin, HE Zhi-Min. Preparation and Properties of Cross-linked Enzyme Aggregates of Papain. Chem. J. Chinese Universities, 2010, 31(9): 1774.

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