TY - JOUR
T1 - Expression of the acidic-subunit of amarantin, carrying the antihypertensive biopeptides VY, in cell suspension cultures of Nicotiana tabacum NT1
AU - Santos-Ballardo, David U.
AU - Germán-Báez, Lourdes J.
AU - Cruz-Mendívil, Abraham
AU - Fuentes-Gutiérrez, Cindy I.
AU - Milán-Carrillo, Jorge
AU - Reyes-Moreno, Cuauhtémoc
AU - Valdez-Ortiz, Angel
N1 - Funding Information:
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 CO-NACYT for the scholarship granted.
PY - 2013/5
Y1 - 2013/5
N2 - A modified version of amarantin, main seed storage protein of Amaranthushypochondriacus, 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.
AB - A modified version of amarantin, main seed storage protein of Amaranthushypochondriacus, 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.
KW - ACE-Inhibitory
KW - Amaranth globulin
KW - Protein hydrolysates
KW - Therapeutic peptides
KW - Transgenic calli
UR - http://www.scopus.com/inward/record.url?scp=84876167980&partnerID=8YFLogxK
U2 - 10.1007/s11240-012-0271-1
DO - 10.1007/s11240-012-0271-1
M3 - Artículo
SN - 0167-6857
VL - 113
SP - 315
EP - 322
JO - Plant Cell, Tissue and Organ Culture
JF - Plant Cell, Tissue and Organ Culture
IS - 2
ER -