title: The acidity of protein fusion partners predominantly determines the efficacy to improve the solubility of the target proteins expressed in Escherichia coli.
abstract: Maximization of the soluble protein expression in Escherichia coli (E. coli) via the fusion expression strategy is usually preferred for academic, industrial and pharmaceutical purposes. In this study, a set of distinct protein fusion partners were comparatively evaluated to promote the soluble expression of two target proteins including the bovine enterokinase largely prone to aggregation and the green fluorescent protein with moderate native solubility. Within protein attributes that are putatively involved in protein solubility, the protein acidity was of particular concern. Our results explicitly indicated the protein fusion partners with a stronger acidity remarkably exhibited a higher capacity to enhance the solubility of the target proteins. Among them, msyB, an E. coli acidic protein that suppresses the mutants lacking function of protein export, was revealed as an excellent protein fusion partner with the distinguished features including high potential to enhance protein solubility, efficient expression, relatively small size and the origin of E. coli itself. In principle, our results confirmed the modified solubility model of Wilkinson-Harrison and especially deepened understanding its essence. Meanwhile, the roles of other parameters such as protein hydrophilicity in solubility enhancement were discussed, a guideline to design or search an optimum protein solubility enhancer was also proposed.
authors: School of Life Sciences, East China Normal University, Zhongshan North Road 3663, Shanghai 200062, China.
authors: Yu
authors: Y
authors: Su
authors: Zhurong
authors: Z
authors: Zou
authors: Shuying
authors: S
authors: Feng
authors: Pei
authors: P
authors: Zhou
authors: Lijuan
authors: L
authors: Cao
mesh_terms: D001709:Biotechnology / Q000379:methods*; D019295:Computational Biology; D004591:Electrophoresis, Polyacrylamide Gel; D004765:Enteropeptidase / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012995:Solubility
chemical_list: D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004765:Enteropeptidase