RESUMEN
Amaranthus hypochondriacus spp. is a commonly grown cereal in Latin America, known for its high protein content. The objective of this study was to separate and identify bioactive peptides found in amaranth seeds through enzymatically-assisted hydrolysis using alcalase and flavourzyme. Hydrolysis was carried out for each enzyme separately and compared to two-step continuous process where both enzymes were combined. The biological activity of the resulting three hydrolysates was analyzed, finding, in general, higher bioactive potential of the hydrolysate obtained in a continuous process (combined enzymes). Its fractions were separated by RP-HPLC, and their bioactivity was analyzed. In particular, two fractions showed the highest biological activity as ACE inhibitors with IC50 at 0.158 and 0.134, thrombin inhibitors with IC50 of 167 and 155, and antioxidants in ABTS assay with SC50 at 1.375 and 0.992 mg/L, respectively. Further sequence analysis of the bioactive peptides was carried out using MALDI-TOF, which identified amino acid chains that have not been reported as bioactive so far. Bibliographic survey allowed identification of similarities between peptides reported in amaranth and other proteins. In conclusion, amaranth proteins are a potential source of peptides with multifunctional activity.
Asunto(s)
Amaranthus/química , Péptidos/química , Proteínas de Plantas/química , Semillas/química , Endopeptidasas/metabolismo , Subtilisinas/metabolismoRESUMEN
Fermentation has shown to be an effective technique in bioactive peptides release. That is why in this study antihypertensive, antithrombotic, and antioxidant activity was evaluated during amaranth proteins fermentation with Lactobacillus casei Shirota and Streptococcus thermophilus 54102 in mono and combined culture. During fermentation an increase of free amine groups was observed, and no statistical differences among monocultures were shown, getting higher concentration in combined culture. This was related to antihypertensive and antioxidant activities, where the highest values were also found in the combined process (45% of angiotensin-converting enzyme inhibition, and 168 µmol Trolox equivalents per liter [TE/L] for 2,2-diphenyl-1-picrylhydrazyl, 268 µmol TE/L for 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid, and 381 µmol Fe2E/L for ferric reducing ability of plasma). On the contrary, antithrombotic activity was not related to free amine groups during fermentation, having the highest bioactivity in different moments in each experiment. L. casei Shirota and S. thermophilus 54102 are strains that are able to release bioactive peptides from amaranth protein, although amaranth is not a common matrix for the development of lactic acid bacteria. In addition, in this study it was observed for the first time that lactic acid strains are able to release bioactive peptides from amaranth protein. In addition, this methodology could be part for the development of fermented beverages, different from fermented milk, to diversify matrix to obtain a novel functional food.
Asunto(s)
Amaranthus/química , Fermentación , Lactobacillales/metabolismo , Péptidos/farmacología , Proteínas de Plantas/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Antihipertensivos/farmacología , Antioxidantes/farmacología , Antitrombinas/farmacología , Ácido Láctico/metabolismo , Lacticaseibacillus casei , Semillas/química , Streptococcus thermophilusRESUMEN
The Cordillera de la Costa is located along the coastline of northern Chile, in the hyperarid Atacama Desert area. Chemical and isotopic analyses of several small coastal springs and groundwater reservoirs between 22.5 °S and 25.5 °S allow understanding groundwater origin, renewal time and the probable timing of recharge. The aquifers are mostly in old volcanic rocks and alluvial deposits. All spring waters are brackish, of the sodium chloride type due to intensive concentration of precipitation due aridity and for deep groundwater to additional water-rock interaction in slowly renewed groundwater and mixing with deep seated brines. The heavy δ18O and δ2H values in spring water are explained by recharge by the arrival of moist air masses from the Pacific Ocean and the originally lighter values in the deep wells can be associated to past recharge by air masses coming from the Atlantic Ocean. Current recharge is assumed almost nil but it was significant in past wetter-than-present periods, increasing groundwater reserves, which are not yet exhausted. To explain the observed chloride content and radiocarbon (14C) activity, a well-mixed (exponential) flow model has been considered for aquifer recharge. The average residence time of groundwater feeding the springs has been estimated between 1 and 2kyr, up to 5kyr and between 7 and 13kyr for deep well water, assuming that current recharge is much less than during the previous wetter period. The recharge period feeding the coastal springs could have been produced 1 to 5kyr BP, when the area was already inhabited, and recharge in the Michilla mine was produced during the 10 to 14.5kyr BP CAPE (Central Andean Pluvial Event) pluvial events of the central Andes. The approximate coincidence of turnover time with the past wet periods, as revealed by paleoclimate data, points to significant recharge during them.