| 海洋真菌Caldariomyces fumago产氯过氧化物酶的双水相提纯条件 |
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| 引用本文： | 陈军,杜宁,许甜甜,夏志华,张书翠,朱旭国.海洋真菌Caldariomyces fumago产氯过氧化物酶的双水相提纯条件[J].生物加工过程,2010,8(6):46-51. |
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| 作者姓名： | 陈军 杜宁 许甜甜 夏志华 张书翠 朱旭国 |
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| 作者单位： | 上海师范大学,生命与环境科学学院,上海,200234 |
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| 基金项目： | 国家自然科学基金资助项目，上海师范大学科技基金资助项目，上海市大学生科研创新活动计划项目 |
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| 摘 要： | 采用聚乙二醇(PEG6000)在(NH4)2SO4高饱和度下沉淀夹带蛋白质富集氯过氧化物酶,再利用磷酸盐溶液复溶解共沉淀物形成的双水相萃取体系高浓度回收酶蛋白,最后再经Sephadex G100柱层析纯化获得高纯度酶试样。结果显示:氯过氧化物酶与PEG共沉淀总活力回收率达85.5%,酶在优化的PEG/磷酸盐双水相系统中上下相分配系数k在0.341以下,酶活力回收率达到69.1%,纯度提高了21.57倍,柱层析可使酶纯度进一步提高到24.79倍,总回收率为37.75%。
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| 关 键 词： | Caldariomyces fumago 氯过氧化物酶 双水相 萃取 |
Purification of chloroperoxidase from a marine fungus Caldariomyces fumago by aqueous two-phase system |
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| CHEN Jun,DU Ning,XU Tian-tian,XIA Zhi-hua,ZHANG Shu-cui,ZHU Xu-guo.Purification of chloroperoxidase from a marine fungus Caldariomyces fumago by aqueous two-phase system[J].Chinese Journal of Bioprocess Engineering,2010,8(6):46-51. |
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| Authors: | CHEN Jun DU Ning XU Tian-tian XIA Zhi-hua ZHANG Shu-cui ZHU Xu-guo |
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| Institution: | (College of Life and Environment Science,Shanghai Normal University,Shanghai 200234,China) |
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| Abstract: | The hydrophilic polymer PEG6000 was used to co-precipitate with chloroperoxidase（CPO）in the saturated ammonium sulfate solution.Then,a aqueous two-phase system of PEG/phosphate was formed based on the co-precipitation resolution to extracted higher composition CPO,and the purity CPO sample from Sephadex G-100 chromatographic system was obtained.Results showed that the CPO yield was 85.5% by co-precipitate with PEG in the presence of the phosphate solution.In the aqueous two-phase system,CPO was dissolved in the bottom phase,the partition coefficient k was lower than 0.341,the yield reached 69.1%,and the purification ratio increased 21.57 times.The purity of CPO was further improved to 24.79 times by column chromatography,the total yield of CPO was 37.75%. |
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| Keywords: | Caldariomyces fumago |
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