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Article

The Fermentation of Orange and Black Currant Juices by the Probiotic Yeast Saccharomyces cerevisiae var. boulardii

by
Andrea Maria Patelski
*,
Urszula Dziekońska-Kubczak
and
Maciej Ditrych
Institute of Fermentation Technology and Microbiology, Faculty of Biotechnology and Food Sciences, Lodz University of Technology, Wolczanska 171/173, 90-530 Lodz, Poland
*
Author to whom correspondence should be addressed.
Appl. Sci. 2024, 14(7), 3009; https://doi.org/10.3390/app14073009
Submission received: 12 March 2024 / Revised: 29 March 2024 / Accepted: 2 April 2024 / Published: 3 April 2024
(This article belongs to the Special Issue Role of Microbes in Agriculture and Food, 2nd Edition)
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Abstract

:
Throughout history, the fermentation of fruit juices has served as a preservation method and has enhanced the retention of bioactive constituents crucial for human well-being. This study examined the possibility of orange and black currant juice fermentation with the probiotic yeast Saccharomyces cerevisiae var. boulardii. Saccharomyces bayanus was used as the reference. The ethanol concentration of the orange juices fermented without added glucose was close to 27 g/L. Adding glucose to the juice increased the alcohol produced by up to 65.58 ± 1.84 g/L (for the orange juice). For the same wort fermented by S. bayanus, the final ethanol concentration was 71.23 ± 1.62 g/L. Regardless of the type of yeast and additives used, the samples retained much of the color of the unfermented juice. The polyphenols content in the fermented samples was close to the initial polyphenols content in the juices. The sensory attributes of the juices fermented by the probiotic yeast did not differ from the samples fermented by S. bayanus. Fermenting fruit juices with probiotic yeasts offers a commercially viable and sensorially appealing method to enhance the product’s value by imparting it with probiotic properties.

1. Introduction

Fermented beverages from ancient times offer essential nutrients crucial for human health, fostering physical and mental well-being while aiding in disease prevention and management. Recently, there has been a surge in consumers’ interest in functional foods that not only provide essential nutrition but also offer specific benefits to bodily functions. Examples include foods rich in bioactive compounds like oligosaccharides, dietary fiber, and beneficial microbes that support gut health. Alongside established functional ingredients such as trace elements, vitamins and probiotics represent a growing category of active components, alongside prebiotics, contributing to overall health and vitality [1].
Live microorganisms that, when administered in adequate quantities, confer health benefits upon the host are called probiotics. Except for lactic acid bacteria, there is significant interest in isolating yeast strains possessing probiotic potential, with Saccharomyces cerevisiae var. boulardii emerging as the yeast species demonstrating clinically significant effects and verified probiotic efficacy. Initially isolated from litchi fruits by French microbiologist Henri Boulard in the 1920s, Saccharomyces cerevisiae var. boulardii (known earlier as Saccharomyces boulardii) has been employed as a probiotic yeast since the 1950s, predominantly within the pharmaceutical industry, often in lyophilized form for the prevention and treatment of diarrhea. Research on commercial varieties of S. cerevisiae var. boulardii revealed morphological and physiological similarities with other Saccharomyces cerevisiae strains, thereby contributing to a deeper understanding of its probiotic characteristics [1,2,3,4]. Multiple investigations have demonstrated the efficacy of S. cerevisiae var. boulardii against diverse enteric pathogens. This effectiveness is attributed to multifaceted mechanisms, such as hindering bacterial adherence and translocation within intestinal epithelial cells, generating substances that counteract bacterial toxins, and the gut cells’ response to pro-inflammatory reactions during bacterial infections [1,4,5].
In addition to its well-known metabolic conversion of sugars into ethanol, yeast manifests other advantageous properties, notably its capacity for phytate biodegradation. This compound is known to form insoluble complexes with essential divalent minerals such as zinc, iron, calcium, and magnesium, which poses challenges by reducing the bioaccessibility of these ions. Moreover, phytate can adversely impact the functional and nutritional characteristics of proteins, including their susceptibility to enzymatic digestion [6].
Another notable function of yeast is folate production. Folates, indispensable cofactors in nucleotide biosynthesis, are vital for cellular proliferation and development. Mammals lack the folate biosynthetic pathway and require an adequate dietary supply, whereas plants and yeast possess this ability. It is well established that folate is important in the prevention of neural tube defects during fetal development. Folate consumption is linked to lowered risks of cardiovascular disease, cancer, and neurodegenerative disorders. Additionally, yeast strains contribute positively to the decomposition of mycotoxins [6,7,8]. The examination of yeast viability demonstrates that probiotic S. cerevisiae var. boulardii withstands the elevated alcohol levels generated during fermentation and subsequent vacuum distillation processes [9,10,11].
Fruit juices contain many compounds, including ascorbic acid, vitamin E, beta-carotene, and phenolic compounds, renowned for their antioxidative properties, which are crucial for neutralizing free radicals. The importance of these antioxidants extends to health maintenance, disease prevention, and age-related degenerative conditions [12,13,14]. The documented protective influence of fruit consumption extends to a spectrum of malignancies affecting the oral cavity, oesophagus, stomach, colorectum, bladder, and other human organs [15]. Orange juice boasts a wealth of bioactive constituents, including ascorbic acid, vitamin B, dietary fiber, potassium, iron, and antioxidant compounds, which are predominantly flavonoids. The quantification of the total phenolic compounds (TPCs) within orange juice and popular nectar formulations from prevalent brands in Brazil exhibited a spectrum spanning from 18.7 to 54.2 mg of gallic acid equivalents (GAE)/100 mL [13].
Black currant (Ribes nigrum) is the runner-up in Europe’s berry cultivation hierarchy. The rich phenolic profile inherent to these berries profoundly influences their sensory characteristics, culminating in pronounced bitterness and astringency. Moreover, the phenolic constituents in black currants hold promise for exerting advantageous effects on human health. Among these compounds, anthocyanins are the principal ones, followed by flavonols, flavan-3-ols, and phenolic acids. The TPC value of black currant juice is close to 40 mg GAE/100 mL [16].
The influence of yeasts on the sensory attributes of finished wine products is crucial and emboldens the significance of yeast selection. Saccharomyces cerevisiae and Saccharomyces bayanus emerge as preeminent choices among wine yeast species due to their resilience to ethanol and adeptness in fermenting high-sugar and acidic juice compositions. The inclusion of probiotic yeasts holds promise in enhancing the beneficial characteristics of fermented foods, owing to the established probiotic attributes of this strain [1,17,18,19,20].
Summing up the abovementioned factors, we evaluated the possibility of orange and black currant juice fermentation with the probiotic yeast S.cerevisiae var. boulardii. The sensory properties of such obtained beverages were also assessed.

2. Materials and Methods

  • Fruit juices
The freshly prepared NFC (not from concentrate) 100% orange or 100% black currant juices from the fruit processing company Tłocznia Szymanowice (Szymanowice, Poland) were used in our experiments.
  • Yeasts and inoculum preparation
The Saccharomyces cerevisiae var. boulardii culture was obtained from the commercial product Enterol 250 (Biocodex, France). Each capsule contained at least 250 mg of lyophilized cells of Saccharomyces cerevisiae var. boulardii CNCM-I-745. The lyophilized yeast was reactivated in 150 mL YPG broth (yeast extract 10 g/L; bactopeptone 20 g/L; glucose 20 g/L; and pH 5.2 ± 0.1) sterilized by autoclaving (121 °C/20 min) and cultivated at 32 ± 1 °C for 24 h with use of a reciprocal laboratory shaker, 140 osc./min. (Eberbach E5900, Belleville, NJ, USA). The post-cultivation suspension containing yeast cells was centrifuged (5000× g, 10 min) and twice washed with a sterile 0.9% NaCl water solution. The final yeast suspension’s dry matter (DM) content was assayed using the spectrophotometer (Rayleight Analytical Instrument, Beijing, China) at 540 nm using the previously prepared standard curve. The second yeast strain—Saccharomyces bayanus BCS103, dry wine strain (Fermentis (Lesaffre), Marquette-lez-Lille, France)—was used to obtain the reference results. One hour before inoculation, the dried yeast cells of this strain were suspended in the sterile 0.9% NaCl solution, and the DM content was assayed for S. cerevisiae var. boulardii. The cells of both strains were used in 0.9% NaCl suspensions at the dose of 250 mg of DM /250 mL of the juice for fermentation.
  • pH and total extract determination
Before the analyses, the samples were centrifuged at 5000× g for 10 min, and the temperature was corrected to 20 ± 1 °C. The pH was measured using an SI Analytics HandyLab 100 pH meter (Xylem Analytics, Mainz, Germany). The total extract was measured using a hand-held digital refractometer (Atago, Tokyo, Japan), and the results were expressed at a Brix degree.
  • Determination of TPC
The total phenolic content was quantified via a spectrophotometric analysis employing the Folin–Ciocalteu’s assay, as detailed by Vieira et al., with slight modifications [21]. In essence, 1.5 mL of either gallic acid aqueous solutions or diluted specific juice was mixed with 2.25 mL of distilled water, followed by the addition of 1.5 mL of Folin–Ciocalteu’s reagent, previously diluted in water (1:10, v/v). This mixture was agitated and left to stand in darkness for 5 min. Subsequently, 1.5 mL of the 6% (w/v) sodium carbonate solution was incorporated, and the reaction was maintained in darkness for 30 min. Following incubation, the absorbance was measured at 725 nm against a blank using a UV9200 UV/Vis spectrophotometer (Rayleight Analytical Instrument, Beijing, China). The results obtained for the gallic acid solutions were used for the standard curve preparation. The blank was prepared by substituting gallic acid or the specific juice with distilled water. The total phenolic content of the orange juice was expressed in milligrams of gallic acid equivalents (GAE) per 100 mL of juice.
  • Juice fermentation
Each fermentation variant was prepared in three replicates. The process was carried out in glass bottles of 1 L capacity filled with 250 mL of the specific fruit juice. The juices were optionally supplemented with diammonium phosphate (DAP), ((NH4)2HPO4) (0.2 g/L of the juice). Before fermentation, the juices were inoculated with a specific yeast solution at the dose 250 mg of DM of cells/L of juice. The bottles were plugged with the fermentation tubes filled with glycerol to limit the oxygen presence in the fermented medium. Alcoholic fermentation was carried out at 29 ± 1 °C for 7 days. After fermentation, the 50 mL samples were separated for chemical analysis and preserved at −20 °C. The remaining volume of the fermented juice was left at 7 ± 1 °C, and after 24 h, the samples were subjected to sensory evaluation.
  • Designation of the samples
The following abbreviated sample designations for the fermented orange juice (OR) and black currant juice (BC) were adopted for ease of sample description: “0”—juice fermented by S. cerevisiae var. boulardii yeast, without the addition of DAP; “+DAP”—juice fermented by the yeast S. cerevisiae var. boulardii with the addition of DAP (0.2 g/L of juice); “+G”—juice fermented by the yeast S. cerevisiae var. boulardii with the addition of glucose (up to a total of glucose and fructose before fermentation equal to 150 g/L of juice); “Sb + G”—juice fermented by the yeast S. bayanus with the addition of DAP (0.2 g/L juice) and glucose (up to a total of glucose and fructose before fermentation equal to 150 g/L juice); “+swe”—juice fermented by the yeast S. cerevisiae var. boulardii with the addition of stevia as the sweetener (4 g/L juice) after fermentation; and “Sb + swe”—juice fermented by S. bayanus yeast with stevia (as the sweetener) added (4 g/L juice) after fermentation.
  • Sensory evaluation
Following a 7-day fermentation period, the samples were allowed to equilibrate for 24 h at 7 ± 1 °C. An in-house trained sensory panel, comprising 12 members from the Department of Biotechnology and Food Sciences at Lodz University of Technology, Poland, conducted the sensory evaluations of the fermented juices. The panelists represented all sexes and diversified age groups (25–65). The panelists were experienced in evaluating fermented beverages by undertaking regular sensory evaluations in similar ongoing research projects. The fermented juice samples were served at approx. 12 °C in aliquots of around 40 mL. In a randomized sequence, the samples were anonymized with a letter code and presented individually to each panelist in single-use plastic cups with lids. Water and unsalted crackers were served between the evaluations to cleanse the palate. The panelists assessed different sensory attributes, including color intensity, clarity, aroma, sweetness, fruitiness, yeastiness, and aftertaste, using a scale ranging from 1 to 10, where 1 represented the lowest intensity and 10 denoted the highest intensity of the specific parameter. A reference mixture comprising the specific juice blended with 5% (v/v) ethanol in the form of 92% rectified spirit was utilized for color, aroma, and fruitiness. The second reference solution was prepared for sweetness by diluting the juice with distilled water at a ratio of 1:10, with a stevia concentration of 6 g/L.
The choice of stevia as a sweetener was made based on the literature data [22,23,24,25,26,27] and our unpublished preliminary studies, the results of which indicated that sweetening fermented juices with stevia, unlike xylitol and erythritol, does not change other sensory characteristics of the product apart from sweetness.
  • Chromatographic analysis of sugars, organic acids, and alcohols
The concentrations of glucose (GLU), fructose (FRU), citric acid (CITacid), malic acid (MALacid), succinic acid (SUCacid), glycerol (GlycOH), and ethanol (EtOH) in the media were analyzed using High-Performance Liquid Chromatography (HPLC) using the Agilent 1260 Infinity system (USA) and a Hi-Plex H+ column (7.7 × 300 mm, 8 µm) (Agilent Technologies, Santa Clara, CA, USA) equipped with a refractive index detector (RID) operating at 55 °C. The column temperature was maintained at 60 °C. A 0.005 M H2SO4 solution of HPLC grade served as the mobile phase, advised by the column manufacturer, flowing at a rate of 0.7 mL/min, and the injected volume of the sample was 20 µL. Prior to the analysis, the samples were mixed with a zinc sulfate solution to achieve its final concentration of 10% for protein precipitation. The sediments were removed through centrifugation at 5000× g for 10 min, and subsequently, the samples were filtered using 0.45 µm PES membranes.
All the assays were carried out in triplicate. The statistical analysis (variance analysis, SD determination, and Student’s t-test at significance level α = 0.05) was performed using the Origin 7.5 computer program.

3. Results and Discussion

3.1. Orange and Black Currant Juice Parameters

The basic parameters of the juices used in our experiments are shown in Table 1.
Before designing the media for fermentation, the basic parameters of the juices (pH, extract, TPC, sugars, and organic acids) were assayed. The pH of the orange juice was equal to 4.18 ± 0.12, while for the black currant juice, this parameter was equal to 3.57 ± 0.11. The pH of the orange juice obtained in our study was a bit higher than the values reported by Cerillo et al. and Escudero-Lopez et al. [28,29] (3.48); Santos et al. [30] (3.7); and Zhang et al. [31] (3.55). The pH value for the black currant juice presented by Kelanne et al. [19] (2.96 ± 0.05) was lower than the result assayed by us. Different orange cultivars, climatic parameters, harvesting time, and methods of juice extraction and preservation may cause these discrepancies. According to the literature, the total extract value of orange juice may vary, depending on cultivar, technology, and other factors, from 11.0 to 13.2 °Brix [13,28,29,30]. While for the orange juice the total extract result obtained by us (11.1 ± 0.2) was consistent with the values reported by others, the Brix degree equal to 7.2 ± 0.1 for the black currant juice obtained by us was only 50% of the values presented in the literature [19,32] (14–16.6 °Brix); in addition to the abovementioned reasons for this difference, the additional elucidation of such a result may be the residual water present in the packaging line that was mixed with the head-part of the pumped juice and thus the extract content in this specific container was lower than expected by the manufacturer.
The total phenolic compound of the orange juice was 49.61 ± 0.63 mg GAE/100 mL, while for the black currant juice, the value of 85.7 ± 0.34 mg GAE/100 mL was noted. According to the literature, the TPC value of the specific fruit juice depends on the cultivar and method of juice extraction. A broad range of TPC values for orange juice, ranging from 18.68 mg GAE/100 mL [13] to 76.8 mg GAE/100 mL, was reported [16,33]. According to the literature, black currant juice may contain as little as 36.7 mg GAE/100 mL [16] and as much as 350 mg GAE/100 mL [32,34]. The values of the TPC obtained in our study fell within this range.
The glucose, fructose, and organic acids presented in Table 1 were determined in the juices using the HPLC technique. The concentration of glucose and fructose in the orange juice was 43.69 ± 1.22 and 47.39 ± 1.07 g/L, respectively; for the blackcurrant juice, the concentration of these sugars was 28.46 ± 0.92 and 30.5 ± 1.03 g/L, respectively. The content of these sugars, crucial for alcoholic fermentation, was similar to the results described by Kelanne et al. [19] for black currant juice (fructose 36.7 and glucose 33.7 g/L) and the results reported by Escudero-Lopez et al. [29] (total reducing sugars 48.5 g/L) and Santos et al. [30] (glucose 57.81 and fructose 56.25 g/L) for orange juice. Zhang et al. [31] reported that the total sugar concentration for orange juice was equal to 70 g/L, which was also consistent with our results presented in Table 1. Organic acids are critical compounds for the flavor bouquet of alcoholic beverages. They are formed due to plant metabolic activity and are also created and utilized by the microflora present during fermentation. The contents of the three acids that we were able to identify using HPLC chromatography presented in Table 1 provide a reference for the concentrations of these acids in the post-fermentation samples in Table 2 and Table 3. The predominant acid in the juices was citric acid, its concentrations being 8.51 ± 0.57 and 5.57 ± 0.23 g/L for the orange and currant juice, respectively. Malic acid (2.56 ± 0.12 g/L) and succinic acid (0.22 ± 0.01 g/L) were also present in the orange juice. The concentration of these two acids was 0.13 ± 0.00 g/L (malic acid) and 0.1 ± 0.00 g/L (succinic acid) for the black currant juice. Kelanne et al. [19] reported the concentration of citric acid in currant juice equal to 31.8 ± 3.4 g/L, a value higher than that obtained in our study. This discrepancy may be due to the different varieties of currant, the time of harvest, the degree of fruit ripeness, and the method of extraction and preservation of the juice. For orange juice, results similar to ours were presented by Zhang et al. [31] (citric acid 3.5 g/L and malic acid 1.5 g/L) and Santos et al. [30] (citric acid 9.72 ± 0.1 g/L; malic acid 4.43 ± 0 g/L; and succinic acid 0.49 g/L). The juices we analyzed before fermentation contained no ethanol or glycerol—Table 1.

3.2. Parameters of the Fermented Juices

The selected organic acids and sugars concentration in the samples after fermentation are shown in Table 2 and Table 3.
After the orange juice fermentation, the samples contained only minor residual fructose (up to 0.73 ± 0.05 g/L for the sample with DAP added), while the glucose concentrations were below the detection levels. These results are indicative of the fully fermented samples. Perhaps even a shorter fermentation time of less than 7 days should be considered.
The predominant organic acid in the juices fermented by S. bayanus and S. cerevisiae var. boulardii yeasts was citric acid. Its concentration in the fermented orange juice ranged from 6.96 ± 0.26 to 7.81 ± 0.24 g/L. The concentration of this acid in the non-fermented orange juice was equal to 8.51 ± 0.57 g/L. Considering the statistics (p < 0.05), it can be concluded that there was a slight reduction in the citric acid concentration only for the OR + DAP and OR “0” samples, while in the case of the glucose-added samples, regardless of the yeast used, the loss of citric acid concentration was not statistically significant (p > 0.05). The malic acid concentration ranged from 1.21 ± 0.10 to 3.12 ± 0.17 g/L, while the succinic acid concentration ranged from 0 to 0.55 ± 0.02 g/L. When we compare these results with the levels of acids in the juices before fermentation, it can be seen that the concentrations of both acids decreased (p < 0.05) for the fermented samples with DAP supplementation and increased (p < 0.05) for the samples with glucose supplementation. In contrast, for the samples fermented without glucose and DAP supplementation (OR “0”), the malic acid and succinic acid concentrations remained at levels similar to those in the unfermented orange juice. Zhang et al. described that, in the case of citric and malic acid in orange juice, there was an increase of about 20% in their concentrations during fermentation by S.cerevisiae SC-125 yeast [31].
The samples after the fermentation of the black currant juice contained small amounts of glucose (up to 1.04 ± 0.06 g/L for the glucose-supplemented juice) and fructose (up to 1.58 ± 0.06 g/L for the glucose-supplemented juice), which may indicate incomplete sugar attenuation due to the too short a processing time or low resistance of the tested strains to the compounds present in black currant juice.
The concentration of citric acid in the fermented currant juice ranged from 4.90 ± 0.27 to 6.75 ± 0.27 g/L. The concentration of this acid in the unfermented currant juice was 5.57 ± 0.23 g/L. It can be concluded that there was a slight increase in the citric acid concentration for the samples fermented with glucose, regardless of the yeast used. The malic acid concentration of the fermented currant juice samples ranged from 0.28 ± 0.01 to 0.31 ± 0.01 g/L, while the succinic acid concentration ranged from 0.13 ± 0.01 to 0.89 ± 0.04 g/L. When we compare these results with the levels of acids in the juices before fermentation, it can be seen that the concentrations of both acids were increased (p < 0.05) for all the fermented samples. Kelanne et al. reported that the concentrations of citric, malic, and succinic acids after alcoholic fermentation of black currant juice carried by the yeasts S.cerevisiae and S. bayanus were in the ranges 30.3–35.1 g/L (for citric acid); 2.6–2.8 g/L (for malic acid); and 0.5–0.9 g/L (for succinic acid) [19].
The ethanol, glycerol, and TPC concentration in the samples after fermentation are shown in Table 4 and Table 5.
The tables above show the results of the ethanol, glycerol, and total phenolic compounds (TPCs) concentrations of the fermented orange (Table 4) and black currant juices (Table 5). The ethanol concentration of the orange juices fermented without added glucose was 27.72 ± 1.28 and 26.45 ± 1.53 g/L for the OR “0” and OR + DAP samples, respectively. In contrast, the glycerol concentration in these samples was 2.42 ± 0.13 and 2.56 ± 0.12 g/L, respectively. These results demonstrate the lack of a positive effect of DAP on the alcohol formation efficiency during orange juice fermentation by the S. cerevisiae var. boulardii strain. Adding glucose to the orange juice undergoing fermentation resulted in a significant increase in the alcohol produced. For the OR + G sample fermented by the S. cerevisiae var. boulardii strain, the ethanol concentration was equal to 65.58 ± 1.84 g/L. In contrast, for the same wort fermented by the S. bayanus yeast, the final ethanol concentration was equal to 71.23 ± 1.62 g/L, meaning that the use of the S. bayanus strain increased the alcoholic fermentation yield of approximately 8.4%, compared to the use of the probiotic yeast strain. Santos et al. [30], for glucose-enriched juices after the fermentation of orange juice by the S. cerevisiae UFLA CA1174 strain, recorded ethanol (58.13 g/L) and glycerol (5.35 g/L) concentrations similar to our results presented in Table 4. Zhang et al. [31] obtained final ethanol concentrations close to 6.5%, which is consistent with our results. In the case of glycerol concentrations, it was observed that replacing the probiotic yeast with the wine strain S. bayanus resulted in a reduction (p < 0.05) in the amount of glycerol produced from 5.58 ± 0.22 g/L to 4.83 ± 0.23 g/L. The TPC values for the fermented orange juices ranged from 43.36 ± 0.42 mg GAE /100 mL (for fermentation with S. bayanus yeast) to 49.81 ± 0.61 mg GAE/100 mL (for the juice without added glucose and DAP fermented with the probiotic strain). Similar TPC values and a minimal increase in this parameter during fermentation (from about 34 to about 35 mg GAE/100 mL) were described by Zhang et al. [31].
For both black currant and orange juice fermentation, the ethanol and glycerol concentrations directly corresponded to the glucose and fructose contents of the samples before fermentation. For the BC “0” sample without supplementation of glucose and DAP to the currant juice, the final ethanol and glycerol concentrations were, respectively, equal to 19.67 ± 1.02 and 2.28 ± 0.12 g/L. Similarly, for the sample with the addition of DAP to the currant juice, the final ethanol and glycerol concentrations were equal to, respectively, 16.14 ± 1.12 and 1.95 ± 0.09 g/L, so it can be concluded that, analogously to the orange juice, no beneficial effect of DAP supplementation was observed on the fermentation carried out by the probiotic yeast S. cerevisiae var. boulardii. Glucose supplementation to the currant juice predictably increased (p < 0.05) the final amounts of ethanol and glycerol produced to 66.05 ± 2.36 and 4.93 ± 0.28 g/L. As with the orange juice, using S. bayanus yeast instead of the probiotic strain induced an increase in the ethanol concentration of about 8% and a decrease in the glycerol concentration of about 5%. Kelanne et al. [19] presented, in their report on the fermentation of currant juices, final ethanol concentrations ranging, depending on the yeast strain used, from 34.6 to 44.7 g/L, which are higher than the values we obtained for the currant juice without added glucose but lower than our results obtained for the juices with added glucose. The TPC values for the fermented currant juices ranged from 81.2 ± 0.41 mg GAE/100 mL (for fermentation with S. bayanus yeast) to 84.1 ± 0.38 mg GAE/100 mL (for the juice without added glucose and DAP fermented with a probiotic strain). Concerning phenolic compounds, Kelanne et al. [19] only reported changes within groups of individual compounds without relating these detailed results to the overall TPC value.

3.3. Sensory Characteristics of the Fermented Juices

The sensory features of the fermented juices are presented in Figure 1 and Figure 2.
The sensory evaluation results of all the juices (Figure 1 and Figure 2) show that, regardless of the type of yeast and additives used, the samples retained much of the color of the unfermented juice, both for the orange and currant juice. The fermented samples with added glucose were slightly sweeter than the fermented juice samples without added glucose. The most noticeable unfavorable yeast aftertaste characterized the samples without added glucose. The yeastiness was positively masked by the addition of 4 g/L of stevia to the samples after fermentation. This sweetener also significantly (p < 0.05) increased the sweetness of the assessed samples. The aftertaste of the evaluated juices was most intense for the samples fermented by probiotic yeast with the addition of stevia after fermentation. The main objective of the sensory evaluation was to compare the juice samples fermented by the probiotic yeast S. cerevisiae var. boulardii with the sensory characteristics of the samples fermented by the wine yeast S. bayanus. The graphs clearly show that the sensory characteristics of the samples differing only in the type of yeast used were very similar—the lines of the graphs overlap in many places. To summarize the sensory evaluation, it can be concluded that using the yeast S. cerevisiae var. boulardii instead of the wine strain S. bayanus does not cause any undesired changes in the taste, aroma, or appearance of the fermented samples. This is positively surprising because probiotic yeast was initially not selected and intended to produce fermented beverages. This conclusion augurs favorably for expanding the use of probiotic yeasts to produce fermented fruit juices with probiotic features. These conclusions are in line with observations on the use of probiotic yeast for the production of fermented beverages presented by de Paula et al., Mulero-Cerezo et al., and Lazo-Velez et al. [9,11,20,35] and also support the statement made by Pinto et al. [36] that fermentation with probiotic yeast offers novel perspectives on optimizing probiotic beverage formulations and indicates opportunities for a holistic approach to enhancing beverage quality.

4. Conclusions

The total polyphenol content is not affected by the fermentation of orange and black currant juices by Saccharomyces cerevisiae var. boulardii. The concentrations of ethanol and glycerol correspond to the contents of glucose and fructose in the samples prior to fermentation. The fermentation of the orange and black currant juice by the probiotic yeast was not affected by DAP supplementation. The use of S. bayanus yeast instead of the probiotic strain induced an increase in the ethanol concentration of about 8% and a decrease in the glycerol concentration of about 5%.
Regardless of the yeast strain and additives used (DAP and glucose), the samples retain much of the color of the unfermented juice. The addition of 4 g/L stevia to the samples after fermentation improved the sensory properties of juices fermented with probiotic yeast. Using the probiotic strain for juice fermentation results in beverages with characteristics and sensory profiles comparable to those produced with the wine strain S. bayanus. Probiotic yeast fermentation appears to be an interesting method for converting fruit juices into value-added products.

Author Contributions

Conceptualization and supervision, A.M.P.; methodology and investigation, A.M.P. and U.D.-K.; software and validation, A.M.P. and M.D.; writing and visualization, A.M.P. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors on request.

Conflicts of Interest

The authors declare no conflicts of interest.

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Applsci 14 03009 g001
Figure 1. Sensory properties of the fermented orange juice.
Figure 1. Sensory properties of the fermented orange juice.
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Figure 2. Sensory properties of the fermented black currant juice.
Figure 2. Sensory properties of the fermented black currant juice.
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Table 1. Parameters of the orange juice and black currant juice used in the experiments.
Table 1. Parameters of the orange juice and black currant juice used in the experiments.
ParameterOrange JuiceBlack Currant Juice
Total extract [°Brix]11.1 ± 0.27.2 ± 0.1
pH [unit]4.18 ± 0.123.57 ± 0.11
GLU [g/L]43.69 ± 1.2228.46 ± 0.92
FRU [g/L]47.39 ± 1.0730.50 ± 1.03
CIT acid [g/L]8.51 ± 0.575.57 ± 0.23
MAL acid [g/L]2.56 ± 0.120.13 ± 0.00
SUC acid [g/L]0.22 ± 0.010.1 ± 0.00
Et OH [g/L]n.d.n.d.
Glyc OH [g/L]n.d.n.d.
TPC [mg GAE/100 mL]49.61 ± 0.6385.7 ± 0.34
n.d.—not detected.
Table 2. Organic acids, glucose, and fructose concentrations in the orange juice after fermentation.
Table 2. Organic acids, glucose, and fructose concentrations in the orange juice after fermentation.
SampleCIT AcidMAL AcidSUC AcidGLUFRU
[g/L]
OR “0”7.05 ± 0.322.50 ± 0.170.18 ± 0.01n.d.n.d.
OR + DAP6.96 ± 0.261.21 ± 0.10n.d.n.d.0.73 ± 0.05
OR + G7.81 ± 0.243.09 ± 0.140.55 ± 0.02n.d.0.17 ± 0.01
OR Sb + G7.62 ± 0.263.12 ± 0.170.45 ± 0.03n.d.0.04 ± 0.01
n.d.—not detected.
Table 3. Organic acids, glucose, and fructose concentrations in the black currant juice after fermentation.
Table 3. Organic acids, glucose, and fructose concentrations in the black currant juice after fermentation.
SampleCIT AcidMAL AcidSUC AcidGLUFRU
[g/L]
BC “0”4.90 ± 0.270.31 ± 0.010.50 ± 0.02n.d.n.d.
BC + DAP5.15 ± 0.280.30 ± 0.010.13 ± 0.010.92 ± 0.06n.d.
BC + G6.04 ± 0.220.30 ± 0.020.71 ± 0.031.04 ± 0.061.58 ± 0.06
BC Sb + G6.75 ± 0.270.28 ± 0.010.89 ± 0.040.74 ± 0.040.95 ± 0.04
n.d.—not detected.
Table 4. Ethanol, glycerol, and TPC content in the orange juice after fermentation.
Table 4. Ethanol, glycerol, and TPC content in the orange juice after fermentation.
SampleEt OHGlyc OHTPC
[g/L]mg/100 mL
OR “0”27.72 ± 1.282.42 ± 0.1349.81 ± 0.61
OR + DAP26.45 ± 1.532.56 ± 0.1248.27 ± 0.53
OR + G65.58 ± 1.845.58 ± 0.2247.81 ± 0.56
OR Sb + G71.23 ± 1.624.83 ± 0.2343.36 ± 0.42
Table 5. Ethanol, glycerol, and TPC content in the black currant juice after fermentation.
Table 5. Ethanol, glycerol, and TPC content in the black currant juice after fermentation.
SampleEt OHGlyc OHTPC
[g/L]mg/100 mL
BC “0”19.67 ± 1.022.28 ± 0.1284.10 ± 0.38
BC + DAP16.14 ± 1.121.95 ± 0.0983.80 ± 0.45
BC + G66.05 ± 2.364.93 ± 0.2883.10 ± 0.36
BC Sb + G72.87 ± 2.424.68 ± 0.2181.20 ± 0.41
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Patelski, A.M.; Dziekońska-Kubczak, U.; Ditrych, M. The Fermentation of Orange and Black Currant Juices by the Probiotic Yeast Saccharomyces cerevisiae var. boulardii. Appl. Sci. 2024, 14, 3009. https://doi.org/10.3390/app14073009

AMA Style

Patelski AM, Dziekońska-Kubczak U, Ditrych M. The Fermentation of Orange and Black Currant Juices by the Probiotic Yeast Saccharomyces cerevisiae var. boulardii. Applied Sciences. 2024; 14(7):3009. https://doi.org/10.3390/app14073009

Chicago/Turabian Style

Patelski, Andrea Maria, Urszula Dziekońska-Kubczak, and Maciej Ditrych. 2024. "The Fermentation of Orange and Black Currant Juices by the Probiotic Yeast Saccharomyces cerevisiae var. boulardii" Applied Sciences 14, no. 7: 3009. https://doi.org/10.3390/app14073009

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