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. 2019 Jul 3;10:1550.
doi: 10.3389/fmicb.2019.01550. eCollection 2019.

Lactic Acid Fermentation of Pomegranate Juice as a Tool to Improve Antioxidant Activity

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Free PMC article

Lactic Acid Fermentation of Pomegranate Juice as a Tool to Improve Antioxidant Activity

E Pontonio et al. Front Microbiol. .
Free PMC article

Abstract

An increasing consumer demand for pomegranate has been globally observed, mainly thanks to the scientific evidence related to its functional and health-promoting features. Pomegranate fruits from twenty accessions identified in Southeastern Italy were characterized according to morphological and chemical features. Juices extracted from pomegranate fruits were fermented with selected Lactobacillus plantarum PU1 and the antioxidant activity investigated. Whey was added to juices to promote the microbial growth. Fermentation led to the increase of the radical scavenging activity (up to 40%) and significant inhibition of the linoleic acid peroxidation. The three fermented juices showing the highest antioxidant activity, and the corresponding unfermented controls, were further characterized. In detail, the cytotoxicity and the protective role toward artificially induced oxidative stress were determined on murine fibroblasts Balb 3T3 through the determination of the viability and the intracellular ROS (reactive oxygen species) scavenging activity (RSA). RSA reached values of ca. 70% in fermented juices, being ca. 40% higher than the unfermented and control samples. Phenols compounds of the pomegranate juices obtained from accessions "Bitonto Piscina," "Sanrà nero," and "Wonderful (reference cultivar) were analyzed through ultrahigh pressure liquid chromatography coupled with mass spectrometry, showing that a marked increase (up to 60%) of the ellagitannins derivatives occurred during fermentation. Sensory analysis showed suitability of the fermented juices to be used as beverage and food ingredient.

Keywords: Balb 3T3; antioxidant activity; ellagitannins; fermentation; lactic acid bacteria; pomegranate.

Figures

FIGURE 1
FIGURE 1
Radical scavenging activity on DPPH (panel A) and ABTS (panel B) of the pomegranate juices prior (Pj) and after (FPj) fermentation with Lactobacillus plantarum PU1. Error bars are shown. a,bDifferent superscript letters referred to significant (P < 0.05) difference between antioxidant activity of the juice prior and after fermentation.
FIGURE 2
FIGURE 2
Kinetics of the linoleic acid peroxidation monitored during 8 days of incubation at 60°C in samples containing the fermented pomegranate juices (FPj) and BHT (1 mg/ml). A blank, corresponding to the reaction mixture without pomegranate juice addition, was included in the analysis.
FIGURE 3
FIGURE 3
Cell viability of mouse fibroblasts treated with freeze-dried pomegranate juices at different concentrations [0.1 mg/ml, panel (A); 1 mg/ml, panel (B); 5 mg/ml, panel (C); and 10 mg/ml, panel (D)]. Viability was determined after 24 (dark gray), 48 (gray), and 72 (light gray) h from the end of the treatment. Data are the means of three independent experiments twice analyzed. Error bars are shown. a-fValues obtained at the same time with different superscript letters differ significantly (P < 0.05).
FIGURE 4
FIGURE 4
Protective effect of different concentrations (1–10 mg/ml) of freeze-dried pomegranate juices and α-tocopherol (α-tp; 250 and 500 μM) on the cell viability of mouse fibroblasts subjected to oxidative stress induced by hydroxide peroxide. The viability of H2O2-stressed cells incubated without antioxidant compounds (reference, rf) was also included. Data are the means of three independent experiments twice analyzed. Error bars are shown. a-cValues obtained at the same concentration with different superscript letters differ significantly (P < 0.05).
FIGURE 5
FIGURE 5
Effect of different concentrations (1–10 mg/ml) of freeze-dried pomegranate juices and α-tocopherol (α-tp; 250 and 500 μM) on the radical scavenging activity (RSA) of mouse fibroblasts subjected to oxidative stress induced by hydroxide peroxide. The RSA of the H2O2-stressed cells incubated without antioxidant compounds (reference, rf) was also included. Data are the means of three independent experiments twice analyzed. Error bars are shown. a-cValues obtained at the same concentration with different superscript letters differ significantly (P < 0.05).
FIGURE 6
FIGURE 6
Sensory analysis of the unfermented [Pj4, Pj18, Pj21, panel (A)] and fermented [FPj4, FPj18, Fpj21, panel (B)] pomegranate juices. Attributes used in the sensory descriptive analysis (Filannino et al., 2013; Di Cagno et al., 2017) are: anise (an), astringent (as), berry (be), fermented (fm), floral (fl), fruity (fr), grape (gr), pungent (pn), sour (sr), sweet (sw), vinegar (vn), wine-like (wn), molasses (ml), color (cl), and browning (br).

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