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Expression of the acidic-subunit of amarantin, carrying the antihypertensive biopeptides VY, in cell suspension cultures of Nicotiana tabacum NT1

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Abstract

A modified version of amarantin, main seed storage protein of Amaranthus hypochondriacus, carrying four antihypertensive biopeptides Val-Tyr into the acidic-subunit of the protein, was expressed in cell suspension cultures of Nicotiana tabacum L. NT1. Cell growth and viability kinetics were assessed to determine optimal conditions for genetic transformation via Agrobacterium tumefaciens. Selection of putative transgenic calli was conducted using 25 μg ml−1 hygromycin. Presence of the transgene was confirmed using histological glucuronidase assay and PCR analysis. Accumulation and expression of the recombinant protein was detected using Western blot analysis. Protein hydrolysate of transgenic calli showed high levels of inhibition of the angiotensin converting enzyme, with an IC50 value of 3.5 μg ml−1. This was 10-fold lower than that of protein extracts of wild-type cells, with an IC50 of 29.0 μg ml−1.

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Acknowledgments

The authors thank to J Castillo-Reyna for providing the biological material and for valuable technical recommendations. This work was supported by grants of Universidad Autónoma de Sinaloa (PROFAPI-UAS, 2011/177). DUSB also thanks to CONACYT for the scholarship granted.

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Authors and Affiliations

  1. Maestría en Ciencia y Tecnología de Alimentos, y Programa Regional de Doctorado en Biotecnología, Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Sinaloa, Av. de las Américas y Josefa Ortiz S/N, 80030, Culiacán, SIN, Mexico

    David U. Santos-Ballardo, Lourdes J. Germán-Báez, Abraham Cruz-Mendívil, Cindy I. Fuentes-Gutiérrez, Jorge Milán-Carrillo, Cuauhtémoc Reyes-Moreno & Angel Valdez-Ortiz

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  1. David U. Santos-Ballardo
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  2. Lourdes J. Germán-Báez
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  3. Abraham Cruz-Mendívil
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  4. Cindy I. Fuentes-Gutiérrez
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  5. Jorge Milán-Carrillo
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  6. Cuauhtémoc Reyes-Moreno
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  7. Angel Valdez-Ortiz
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Correspondence to Angel Valdez-Ortiz.

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Santos-Ballardo, D.U., Germán-Báez, L.J., Cruz-Mendívil, A. et al. Expression of the acidic-subunit of amarantin, carrying the antihypertensive biopeptides VY, in cell suspension cultures of Nicotiana tabacum NT1. Plant Cell Tiss Organ Cult 113, 315–322 (2013). https://doi.org/10.1007/s11240-012-0271-1

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