论著

一种新型尿酸氧化酶四聚体提取纯化方法的建立

  • 江筠 张林焱 林慧娟 郝倩雯 田石华 秦荣浦 刘敏
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  • 上海生物制品研究所有限责任公司第2研究室 200051

网络出版日期: 2025-08-16

Establishment of a new method for extraction and purification of urate oxidase tetramer

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  • No. 2 Research Laboratory, Shanghai Institute of Biological Products Co., Ltd., Shanghai 200051, China

Online published: 2025-08-16

摘要

目的 建立一种简便、快速的尿酸氧化酶四聚体提取纯化的方法,以适应大规模生产需求。方法 首先通过基因工程的方法获得可以稳定发酵生产尿酸氧化酶的大肠埃希菌菌株,再通过一步萃取法从破碎菌体沉淀中获得尿酸氧化酶蛋白混合物,通过硫酸铵沉淀、离子交换和分子筛层析,最终获得纯化的活性尿酸氧化酶四聚体。结果 通过优化发酵及提取条件,每升发酵液可得到尿酸氧化酶四聚体蛋白约400 mg,其纯度>95%,比活>10 U/mg。结论 建立了一种新的尿酸氧化酶四聚体高效提取纯化的方法。

本文引用格式

江筠 张林焱 林慧娟 郝倩雯 田石华 秦荣浦 刘敏 . 一种新型尿酸氧化酶四聚体提取纯化方法的建立[J]. 国际生物制品学杂志, 2019 , 42(3) : 116 -119 . DOI: 10.3760/cma.j.issn.1673-4211.2019.03.004

Abstract

Objective  To establish a simple and fast method for extraction and purification of urate oxidase tetramer. Methods  Genetic engineering was used to obtain an E. coli strain which can produce urate oxidase effectively. The mixture of urate oxidase was obtained from precipitant of bacteria lysis. The purified tetramer of urate oxidase was extracted by ammonium sulfate precipitation, ion exchange chromatography and molecular sieve chromatography. Results  Approximately 400 mg tetramer protein was obtained from each liter of fermentation with purity >95% and specific activity >10 U/mg. Conclusion  An efficient method for extraction and purification of urate oxidase tetramer is established.
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