Development of a protein-solubilizing expression method based on the synergistic action of intein ΔI-CM and the solubility enhancer elastin-like polypeptide

Rai, Kamal; Chu, Xiaohui; Zhou, Dongyan; Li, Fēi; Yang, Junqing; Lin, Jingtao; Shen, Shijun; Song, Haipeng; Sun, Yue; Nian, Rui

doi:10.1016/j.bej.2020.107900

Cited by 3 publications

(1 citation statement)

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“…Mini-Intein ∆I-CM, an intronic peptide with self-cleaving activity at the C-terminal only, was developed by Wood et al 23 . Then Sun et al established a novel soluble protein expression system based on the synergistic interaction of elastin-like polypeptide (ELP) and Mini-Intein ∆I-CM, the fusion of which allowed the target protein to be directly separated from ELP in a mild and enzyme-free environment, allowing for more e cient isolation and puri cation of the target protein 24 .…”

Section: Introductionmentioning

confidence: 99%

Engineering self-cleavage fusion system for the Production of Chimera Spider Silk Proteins

zhou,

Huang,

et al. 2024

Preprint

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The spider silk protein (spidroin), which has powerful mechanical properties, has been extensively studied and shown potential application in various fields. The predatory nature of spiders makes native spidroin challenging to obtain, while heterologous expression of spidroin was hindered by the gene sequence features such as highly repetitive regions and high GC content. The low yield of spidroin subsequently affects its further application. In this study, we constructed a convenient expression system by employing a fusion tag in combination with a self-cleavage intronic peptide (intein) for three kinds of chimeric spidroins with different numbers of repetitive units, and soluble expression of the three kinds of spidroins after optimizing expression conditions was achieved with yields of 266 mg/L (NT2RepCT), 135 mg/L (NT4RepCT), and 125 mg/L (NT6RepCT), respectively. Three kinds of chimeric spidroins displayed increased β-sheet content with increased repetitive units during the transition from the solution to the dry state. Their capacity to form filamentous fibrils increased with the number of repetitive numbers. This study provides a solution for spidroin soluble expression and lays a foundation for its future application.

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Section: Introductionmentioning

confidence: 99%