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Bioimpacts. 2020;10(4): 259-268.
doi: 10.34172/bi.2020.33

PMCID: PMC7502910
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Original Research

Designing a new generation of expression toolkits for engineering of green microalgae; robust production of human interleukin-2

Jaber Dehghani 1,2 ORCID logo, Khosro Adibkia 3* ORCID logo, Ali Movafeghi 2, Mohammad M. Pourseif 1, Yadollah Omidi 1,3,4* ORCID logo

1 Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran
2 Department of Plant Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran
3 Department of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
4 Department of Pharmaceutical Sciences, College of Pharmacy, Nova Southeastern University, Fort Lauderdale, Florida, USA
*Corresponding authors: Khosro Adibkia, Email: ; Yadollah Omidi, Email: yomidi@tbzmed.ac.ir Adibkia@tbzmed.ac.ir

Abstract

Introduction: Attributable to some critical features especially the similarity of the protein synthesis machinery between humans and microalgae, these microorganisms can be utilized for the expression of many recombinant proteins. However, low and unstable gene expression levels prevent the further development of microalgae biotechnology towards protein production.
Methods: Here, we designed a novel “Gained Agrobacterium-2A plasmid for microalgae expression” (named GAME plasmid) for the production of the human interleukin-2 using three model microalgae, including Chlamydomonas reinhardtii, Chlorella vulgaris, and Dunaliella salina. The GAME plasmid harbors a native chimeric hsp70/Int-1/rbcS2 promoter, the microalgae specific Kozak sequence, a novel hybrid 2A peptide, and Int-1 and Int-3 of the rbcS2 gene in its expression cassette.
Results: The obtained data confirmed that the GAME plasmid can transform the microalgae with high transformation frequency. Molecular and proteomic analyses revealed the stable and robust production of the hIL-2 by the GAME plasmid in the microalgae. According to the densimetric analysis, the microalgae can accumulate the produced protein about 0.94% of the total soluble protein content. The ELISA data confirmed that the produced hIL-2 possesses the same conformation pattern with the acceptable biological activity found naturally in humans.
Conclusion: Most therapeutic proteins need post-translational modifications for their correct conformation, biological function, and half-life. Accordingly, microalgae could be considered as a cost-effective and more powerful platform for the production of a wide range of recombinant proteins such as antibodies, enzymes, hormones, and vaccines.
Keywords: 2A peptide, Agrobacterium tumefaciens, Chlamydomonas reinhardtii, Chlorella vulgaris, Dunaliella salina, Interleukin-2
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Submitted: 01 Jun 2020
Revision: 17 Jun 2020
Accepted: 29 Jun 2020
ePublished: 13 Jul 2020
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