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The effects of AtEXO70E2 overexpression on the secretion and production of recombinant proteins in tobacco BY-2 suspension cells

  • Molecular Pharming/Molecular Farming
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In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

As a host for recombinant protein expression, plants have their own advantages over microorganisms and animal cells. When plant cells are used to express recombinant proteins, the extracellular secretion of recombinant proteins can simplify the purification process thereby reducing production costs. In view of the low efficiency of secretion mediated by traditional secretion signal peptides, this study attempted to increase the secretion rate by using the exocyst-positive organelle (EXPO) in the non-traditional secretion pathway. The results showed that overexpression of the Arabidopsis EXO70E2 gene could increase the secretion rate and production of enhanced green fluorescent protein located in the endoplasmic reticulum (ER) or secretory pathway but had no significant effect on the one located in the cytoplasm. In order to increase the effect of EXO70E2, this study further optimized the culture conditions of stable transformed cell lines through design of experiment. The verification on two recombinant proteins, IL10 and trastuzumab, confirmed that the co-expression of EXO70E2 combined with the optimization of culture conditions can increase the secretion rate and yield of these recombinant proteins, providing a new strategy for the development of plant bioreactors.

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Data Availability

Data available on request from the authors.

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Funding

This work was funded by the National Natural Science Foundation of China (31800256) and the Natural Science Foundation of Anhui Province (2108085MC84).

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Authors and Affiliations

  1. Department of Bioengineering, Huainan Normal University, Huainan, 232038, Anhui Province, China

    Ya Li, Juan Han, Yugu Xu, Yun Wang & Guopeng Miao

  2. Key Laboratory of Bioresource and Environmental Biotechnology of Anhui Higher Education Institutes, Huainan Normal University, Huainan, 232038, Anhui Province, China

    Ya Li, Yun Wang & Guopeng Miao

  3. Leafyland Biopharma Co., Ltd., Jiaxing, Zhejiang, 314000, Province, China

    Guopeng Miao

Authors
  1. Ya Li
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  2. Juan Han
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  3. Yugu Xu
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  4. Yun Wang
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  5. Guopeng Miao
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Contributions

G.M. designed research; Y.L., J.H., and Y.X. performed research; G.M. and Y.W. analyzed data; Y.W. contributed new reagents/analytic tools; and G.M. wrote the paper.

Corresponding author

Correspondence to Guopeng Miao.

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The authors declare no competing interests.

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Li, Y., Han, J., Xu, Y. et al. The effects of AtEXO70E2 overexpression on the secretion and production of recombinant proteins in tobacco BY-2 suspension cells. In Vitro Cell.Dev.Biol.-Plant 59, 692–699 (2023). https://doi.org/10.1007/s11627-023-10379-9

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  • DOI: https://doi.org/10.1007/s11627-023-10379-9

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