| 木瓜蛋白酶交联聚体的制备及性质 |
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| 引用本文： | 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[J]. 高等学校化学学报, 2010, 31(9): 1774 |
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| 作者姓名： | 王梦凡 齐崴 苏荣欣 何志敏 |
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| 作者单位： | 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 |
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| 基金项目： | 国家自然科学基金，国家"八六三"计划项目，教育部新世纪优秀人才项目，教育部科学技术研究重点项目，天津市应用基础研究计划项目 |
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| 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.
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| 关 键 词： | 交联酶聚体 固定化 木瓜蛋白酶 稳定性 形貌 集簇 |
| 收稿时间： | 2009-12-14 |
Preparation and Properties of Cross-linked Enzyme Aggregates of Papain |
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| WANG Meng-Fan,QI Wei,SU Rong-Xin,HE Zhi-Min. Preparation and Properties of Cross-linked Enzyme Aggregates of Papain[J]. Chemical Research In Chinese Universities, 2010, 31(9): 1774 |
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| Authors: | WANG Meng-Fan QI Wei SU Rong-Xin HE Zhi-Min |
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| Affiliation: | State Key Laboratory of Chemical Engineering(Tianjin University), Chemical Engineering Research Center, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China |
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| 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. |
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| Keywords: | Cross-linked enzyme aggregate Immobilization Papain Stability Morphology Cluster |
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