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Open AccessArticle

Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m

1
Institute for Multidisciplinary Research in Applied Biology, UPNA, 31006 Pamplona, Spain
2
National Centre for Biotechnology, Plant Molecular Genetics Department, CSIC, 28049 Madrid, Spain
3
Hepatology Program, University of Navarra, CIMA, E-31008 Pamplona, Spain
4
CIBERehd, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, Spain
*
Author to whom correspondence should be addressed.
Both authors contributed equally to this work.
Plants 2020, 9(2), 183; https://doi.org/10.3390/plants9020183
Received: 5 December 2019 / Revised: 24 January 2020 / Accepted: 30 January 2020 / Published: 2 February 2020
(This article belongs to the Special Issue Plant Molecular Farming)
Human cardiotrophin 1 (CT1), a cytokine with excellent therapeutic potential, was previously expressed in tobacco chloroplasts. However, the growth conditions required to reach the highest expression levels resulted in an impairment of its bioactivity. In the present study, we have examined new strategies to modulate the expression of this recombinant protein in chloroplasts so as to enhance its production and bioactivity. In particular, we assessed the effect of both the fusion and co-expression of Trx m with CT1 on the production of a functional CT1 by using plastid transformation. Our data revealed that the Trx m fusion strategy was useful to increase the expression levels of CT1 inside the chloroplasts, although CT1 bioactivity was significantly impaired, and this was likely due to steric hindrance between both proteins. By contrast, the expression of functional CT1 was increased when co-expressed with Trx m, because we demonstrated that recombinant CT1 was functionally active during an in vitro signaling assay. While Trx m/CT1 co-expression did not increase the amount of CT1 in young leaves, our results revealed an increase in CT1 protein stability as the leaves aged in this genotype, which also improved the recombinant protein’s overall production. This strategy might be useful to produce other functional biopharmaceuticals in chloroplasts. View Full-Text
Keywords: cardiotrophin-1; thioredoxin; plastid transformation; bioactivity; tobacco cardiotrophin-1; thioredoxin; plastid transformation; bioactivity; tobacco
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MDPI and ACS Style

Ancín, M.; Sanz-Barrio, R.; Santamaría, E.; Fernández-San Millán, A.; Larraya, L.; Veramendi, J.; Farran, I. Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m. Plants 2020, 9, 183. https://doi.org/10.3390/plants9020183

AMA Style

Ancín M, Sanz-Barrio R, Santamaría E, Fernández-San Millán A, Larraya L, Veramendi J, Farran I. Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m. Plants. 2020; 9(2):183. https://doi.org/10.3390/plants9020183

Chicago/Turabian Style

Ancín, María; Sanz-Barrio, Ruth; Santamaría, Eva; Fernández-San Millán, Alicia; Larraya, Luis; Veramendi, Jon; Farran, Inmaculada. 2020. "Functional Improvement of Human Cardiotrophin 1 Produced in Tobacco Chloroplasts by Co-Expression with Plastid Thioredoxin m" Plants 9, no. 2: 183. https://doi.org/10.3390/plants9020183

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