| 超滤膜分离纯化珊瑚藻溴过氧化物酶 |
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| 引用本文： | 程小飞,章表明,薛松,肖通虎.超滤膜分离纯化珊瑚藻溴过氧化物酶[J].中国生物工程杂志,2011,31(3):76-80. |
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| 作者姓名： | 程小飞 章表明 薛松 肖通虎 |
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| 作者单位： | 1. 宁波大学材料科学与化学工程学院 宁波 315211;
2. 中国科学院大连化学物理研究所 大连 116023;
3. 中国科学院研究生院 北京 100049 |
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| 基金项目： | 国家“973”计划(2009CB724700); 中国科学院知识创新工程重要方向项目(KSCX2-YW-G-073); 浙江省自然科学基金(Y407353); 浙江省公益技术研究项目(2010C31099)资助项目 |
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| 摘 要： | 利用超滤膜对珊瑚藻中溴过氧化物酶(EC 1.11.1.18,分子质量740kDa)进行分离纯化,对膜的截留分子质量、操作压力、起始蛋白质浓度、搅拌速率、pH、离子强度等条件进行了优化。超滤分离纯化最优条件为:截留分子质量为100kDa的聚偏氟乙烯(PVDF)膜,操作压力为0.02MPa,搅拌速率为600r/min,起始蛋白质浓度为0.1g/L,pH=6.0。对粗酶液先进行热沉淀纯化,再进行超滤纯化,溴过氧化物酶被纯化了21倍,比活为212U/mg,酶活回收率为96%。
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| 关 键 词： | 聚偏氟乙烯(PVDF)膜 超滤 纯化 溴过氧化物酶 |
| 收稿时间： | 2010-11-19 |
| 修稿时间： | 2011-01-13 |
Isolation and Purification of Bromoperoxidase from Corallina Officinalis Using Ultrafiltration Membrane |
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| CHENG Xiao-fei,ZHANG Biao-ming,XUE Song,XIAO Tong-hu.Isolation and Purification of Bromoperoxidase from Corallina Officinalis Using Ultrafiltration Membrane[J].China Biotechnology,2011,31(3):76-80. |
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| Authors: | CHENG Xiao-fei ZHANG Biao-ming XUE Song XIAO Tong-hu |
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| Institution: | CHENG Xiao-fei1 ZHANG Biao-ming2,3 XUE Song2 XIAO Tong-hu1(1 Faculty of Materials Science and Chemical Engineering,Ningbo University,Ningbo 315211,China)(2 Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,China)(3 Graduate University of Chinese Academy of Sciences,Beijing 100049,China) |
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| Abstract: | A novel, simple and highly efficient process for purifying vanadium bromoperoxidase from Corallina officinalis is developed, which is consist of two steps: crude enzyme extraction was heated at 70℃ for 2h, and then was ultrafiltered with PVDF membrane. The ultrafiltration conditions were optimized, and the optimized conditions are: MWCO=100kDa, 0.02MPa pressure, 600r/min stirring, 0.12mg/ml initial protein concentration and pH=6.0. The crude extraction purified with heat treatment and then ultrafitration results a 21-fold purification, with a 96% overall yield and 212U/mg specific activity. |
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| Keywords: | Vanadium Bromoperoxidase Polyvinylidenefluoride(PVDF) membrane Ultrafiltration Purification |
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