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Application of short hydrophobic elastin-like polypeptides for expression and purification of active proteins

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Abstract

To investigate the application of short elastin-like polypeptides (ELPs) in the purification of bioactive proteins, short hydrophobic ELP[I]n (n = 30, 40, 50) tags were constructed. Both the ELP[I]n tags and the ELP[I]n–Trx fusion proteins could be stably expressed in Escherichia coli and purified by inverse transition cycling, respectively. Total protein concentrations determined by BCA protein assay showed that the yield of the fusion proteins decreased with increasing ELP length. Measurements of the inverse transition temperature (Tt) of the ELP[I]n–Trx under different salts or PEG8000 concentrations showed decreased Tt upon elevated concentrations; while, all the Tts were suitable for generating proteins from 4 to 37.5 ºC. Furthermore, to identify a linker peptide for bioactive protein production without the need to remove the ELP[I]n tag, the activity of eGFP protein fused with ELP[I]30 tag by either a poly-N or a G4S linker was quantified using a fluorescence spectrophotometer. The results indicated that the ELP[I]30–eGFP fusion proteins with the poly-N linker showed higher fluorescence levels than those with the G4S linker. Our results demonstrated that short ELP[I]n tags with low Tt were useful in protein expression and purification, and poly-N linker played the key role in producing bioactive proteins without the need to remove the ELPs.

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Acknowledgements

This study was supported by grants from the National Natural Science Foundation of China (Grant No. 31370937 to Xue-Jun Hu; Grant No. 21505013 to Jing Sun) and the Dalian Science and Technology Innovation Funds (Grant No. 2018J13SN087 to Ming-Fei Lang).

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Authors and Affiliations

  1. Medical College, Dalian University, Dalian, 116622, Liaoning, China

    Chun-Guang Yang, Ming-Fei Lang, Xin Fu, Heng Lin, Li-Chao Zhang, Gao-Shun Ge & Xue-Jun Hu

  2. College of Environment and Chemical Engineering, Dalian University, Dalian, 116622, Liaoning, China

    Jing Sun

Authors
  1. Chun-Guang Yang
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  2. Ming-Fei Lang
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  3. Xin Fu
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  4. Heng Lin
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  5. Li-Chao Zhang
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  6. Gao-Shun Ge
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  7. Jing Sun
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  8. Xue-Jun Hu
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Contributions

X-JH and JS conceived and designed the study. C-GY, M-FL, XF, HL, L-CZ, and G-SG carried out the experiment. C-GY and M-FL co-wrote the manuscript. All authors analyzed the results and corrected the manuscript.

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Correspondence to Jing Sun or Xue-Jun Hu.

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The authors declared that they have no conflicts of interest to this work.

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Yang, CG., Lang, MF., Fu, X. et al. Application of short hydrophobic elastin-like polypeptides for expression and purification of active proteins. 3 Biotech 10, 156 (2020). https://doi.org/10.1007/s13205-020-2139-0

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