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首页> 外文期刊>BMC Biotechnology >Ubiquitin-like prokaryotic MoaD as a fusion tag for expression of heterologous proteins in Escherichia coli
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Ubiquitin-like prokaryotic MoaD as a fusion tag for expression of heterologous proteins in Escherichia coli

机译：泛素样原核生物MoaD作为在大肠杆菌中表达异源蛋白的融合标签

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Background Eukaryotic ubiquitin and SUMO are frequently used as tags to enhance the fusion protein expression in microbial host. They increase the solubility and stability, and protect the peptides from proteolytic degradation due to their stable and highly conserved structures. Few of prokaryotic ubiquitin-like proteins was used as fusion tags except ThiS, which enhances the fusion expression, however, reduces the solubility and stability of the expressed peptides in E. coli . Hence, we investigated if MoaD, a conserved small sulfur carrier in prokaryotes with the similar structure of ubiquitin, could also be used as fusion tag in heterologous expression in E. coli . Results Fusion of MoaD to either end of EGFP enhanced the expression yield of EGFP with a similar efficacy of ThiS. However, the major parts of the fusion proteins were expressed in the aggregated form, which was associated with the retarded folding of EGFP, similar to ThiS fusions. Fusion of MoaD to insulin chain A or B did not boost their expression as efficiently as ThiS tag did, probably due to a less efficient aggregation of products. Interestingly, fusion of MoaD to the murine ribonuclease inhibitor enhanced protein expression by completely protecting the protein from intracellular degradation in contrast to ThiS fusion, which enhanced degradation of this unstable protein when expressed in E. coli . Conclusions Prokaryotic ubiquitin-like protein MoaD can act as a fusion tag to promote the fusion expression with varying mechanisms, which enriches the arsenal of fusion tags in the category of insoluble expression.
机译：背景真核遍在蛋白和SUMO经常用作增强微生物宿主中融合蛋白表达的标签。它们增加了溶解度和稳定性，并由于其稳定且高度保守的结构而保护了肽免于蛋白水解降解。除ThiS外，很少使用原核泛素样蛋白作为融合标签，ThiS可增强融合表达，但会降低表达的肽在大肠杆菌中的溶解度和稳定性。因此，我们研究了MoaD（原核生物中一种保守的小硫载体，具有与泛素相似的结构）是否也可以用作大肠杆菌中异源表达的融合标签。结果MoaD融合到EGFP的任一端提高了EGFP的表达产量，具有与ThiS相似的功效。但是，融合蛋白的主要部分以聚集形式表达，这与EGFP的折叠延迟有关，类似于ThiS融合。 MoaD与胰岛素链A或B的融合不能像ThiS标签那样有效地增强其表达，这可能是由于产物聚集效率较低。有趣的是，与ThiS融合相反，MoaD与鼠核糖核酸酶抑制剂的融合通过完全保护蛋白质免受细胞内降解而增强了蛋白质表达，而ThiS融合则在大肠杆菌中表达时增强了该不稳定蛋白质的降解。结论原核泛素样蛋白MoaD可作为融合标签，以不同的机制促进融合表达，丰富了融合标签中不溶性表达的库。

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