Expression and purification of lacticin Q by small ubiquitin-related modifier fusion in Escherichia coli
Expression and purification of lacticin Q by small ubiquitin-related modifier fusion in Escherichia coli
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DOI：
10.1007/s12275-012-1425-x
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发表时间：
2012-04
期刊：
影响因子：
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通讯作者：
Qingshan Ma;Zhanqiao Yu;B. Han;Qing Wang;Rijun Zhang
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文献类型：
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作者：
Qingshan Ma;Zhanqiao Yu;B. Han;Qing Wang;Rijun Zhang
乳酸菌素Q是一种广谱的II类细菌素，具有替代传统抗生素的潜力。本研究的目的是利用一个小的泛素相关修饰物(SUMO)融合蛋白表达系统生产重组乳糖素Q。将168bp的lacticin Q基因克隆到表达载体pET SUMO中，转化大肠杆菌BL21(DE3)。经Ni-NTA琼脂糖柱纯化，每升发酵液可获得130 mg的融合蛋白。然后将相扑标签从蛋白质中切割出来，并将其重新应用到柱子上。最终得到约32 mg的乳酸菌素Q(≥96%纯度)。该重组蛋白具有与天然蛋白相似的抗菌特性，表明该融合系统已成功表达了Lacticin Q。
Lacticin Q is a broad-spectrum class II bacteriocin with potential as an alternative to conventional antibiotics. The objective of this study was to produce recombinant lacticin Q using a small ubiquitin-related modifier (SUMO) fusion protein expression system. The 168-bp lacticin Q gene was cloned into the expression vector pET SUMO and transformed intoEscherichia coliBL21(DE3). The soluble fusion protein was recovered with a Ni-NTA Sepharose column (95% purity); 130 mg protein was obtained per liter of fermentation culture. The SUMO tag was then proteolytically cleaved from the protein, which was re-applied to the column. Finally, about 32 mg lacticin Q (≥96% purity) was obtained. The recombinant protein exhibited antimicrobial properties similar to that of the native protein, demonstrating that lacticin Q had been successfully expressed by the SUMO fusion system.