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在大肠杆菌中通过小泛素相关修饰物融合生产细菌素 E50-52。

Production of bacteriocin E50-52 by small ubiquitin-related modifier fusion in Escherichia coli.

出版信息

Pol J Microbiol. 2013;62(4):345-50.

Abstract

Bacteriocin E50-52, a class IIa bacteriocin with a wide antibacterial spectrum, and has a huge potential to be a substitute for convention. antibiotics. In this research, the bacteriocin E50-52 gene was cloned into the expression vector pET SUMO (small ubiquitin-related modifier) and introduced into Escherichia coli BL21 (DE3). The recombinant fusion protein SUMO-bacteriocin E50-52 expressed in a soluble form was purified to a purity of more than 90% by Ni-NTA sepharose column and 117 mg fusion protein was obtained per liter of fermentation culture. The fusion protein was cleaved with SUMO protease and re-applied to a Ni-NTA Sepharose column. Finally, about 16 mg recombinant bacteriocin E50-52 (rbE50-52) was obtained from a 1-liter fermentation culture with no less than 95% purity. The rbE50-52 had similar antimicrobial properties and molecular weight as the native bacteriocin E50-52 and showed very low hemolytic activity.

摘要

细菌素 E50-52 是一种广谱抗菌的 IIa 类细菌素,具有巨大的替代传统抗生素的潜力。在这项研究中,细菌素 E50-52 基因被克隆到表达载体 pET SUMO(小泛素相关修饰物)中,并引入大肠杆菌 BL21(DE3)。重组融合蛋白 SUMO-细菌素 E50-52 以可溶形式表达,通过 Ni-NTA sepharose 柱纯化至纯度超过 90%,每升发酵培养物可获得 117 毫克融合蛋白。融合蛋白用 SUMO 蛋白酶切割,然后重新应用于 Ni-NTA Sepharose 柱。最后,从 1 升发酵培养物中获得了约 16 毫克重组细菌素 E50-52(rbE50-52),纯度不低于 95%。rbE50-52 具有与天然细菌素 E50-52 相似的抗菌特性和分子量,且溶血活性非常低。

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