Rapid and efficient one-step purification of a serralysin family protease by using a p-aminobenzamidine-modified affinity medium

文献类型： 外文期刊

第一作者： Li, Shangyong

作者： Li, Shangyong;Wang, Linna;Yang, Juan;Ma, Zibin;Wang, Yuejun;Liu, Junzhong;Hao, Jianhua;Sun, Mi;Hao, Jianhua;Sun, Mi;Lin, Shengxiang;Lin, Shengxiang

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关键词： affinity purification;p-aminobenzamidine;serralysin family proteases

期刊名称：JOURNAL OF SEPARATION SCIENCE （ 影响因子：3.645； 五年影响因子：2.943 ）

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收录情况： SCI

摘要： The metalloproteinase MP belongs to the serralysin family, which is involved in important functions such as nutrient acquisition and infection pathogenesis. Serralysin proteases in highly purified form are commonly used at the industrial level with several purposes. In this study, we set up an efficient and rapid purification protocol for MP using a p-aminobenzamidine-modified affinity chromatography. The affinity medium was synthesized by using p-aminobenzamidine as affinity ligand immobilized via cyanuric chloride spacer to Sepharose 6B sorbent carrier. According to the adsorption analysis, the dissociation constant K-d and theoretical maximum adsorption Q(max) of this medium were 24.2 mu g/mL and 24.1 mg/g wet sorbent, respectively. The purity of MP was assessed by a high-performance liquid chromatography on a TSK3000SW column and sodium dodecyl sulfate polyacrylamide gel electrophoresis, revealing values of 98.7 and similar to 98%, respectively. The specific activity of purified MP was 95.6 U/mg, which is similar to values obtained through traditional purification protocols. In conclusion, our protocol could be easily employed for the rapid isolation of MP with high purity, and could be implemented for other serralysin family proteases.

分类号： O6

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[1]Affinity purification of metalloprotease from marine bacterium using immobilized metal affinity chromatography. Li, Shangyong,Wang, Linna,Yang, Juan,Bao, Jing,Liu, Junzhong,Hao, Jianhua,Sun, Mi,Li, Shangyong,Wang, Linna,Yang, Juan,Bao, Jing,Liu, Junzhong,Hao, Jianhua,Sun, Mi,Li, Shangyong,Wang, Linna,Yang, Juan,Bao, Jing,Liu, Junzhong,Hao, Jianhua,Sun, Mi,Lin, Shengxiang.

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