Mixed Starter Research Articles

Biotechnological potential of non-Saccharomyces yeasts isolated during spontaneous fermentations of Malvar (Vitis vinifera cv. L.)

Non-Saccharomyces yeast species assume an important role in wine flavor. Notwithstanding, the chemical basis for the flavor characteristics of wines from some grape varieties is not yet defined. The value of this work lies in the use of Malvar white grape, an autochthonous variety from Madrid (Spain) winegrowing region to conduct spontaneous fermentations. This is the first time that a comparative characterization of a wide range of non-Saccharomyces species and a comprehensive analysis of these yeast-derived volatiles has been carried out in this grape variety. β-glucosidase and pectinase (polygalacturonase) extracellular activities were tested on agar plates as primary selection criteria among the 504 non-Saccharomyces isolated from Malvar spontaneous fermentations during four consecutive harvests. Analysis of the wines obtained after fermentation using the selected yeast strains indicates that non-Saccharomyces yeasts isolated along the fermentative process seem that could have a positive impact, showing a high variability in the volatile compounds contributing to the organoleptic characteristics of Malvar wines. Torulaspora delbrueckii CLI 918 was defined as the yeast strain with potential interest for its contribution to the aromatic wine profile with flowery and fruity aromas and could be used in mixed starter cultures with Saccharomyces cerevisiae. However, Hanseniaspora guilliermondii increased the volatile acidity and ethyl acetate, but this species along with the genus Pichia and Candida seem to provide a high quantity of extracellular enzymes which may be beneficial in wine making.

Evaluation of Williopsis saturnus Inoculum Level on Fermentation and Flavor Compounds of White Wines Made from Emir ( Vitis vinifera L.) Grown in Anatolia

The effects of different W. saturnus inoculum levels together with S. cerevisiae on fermentation and production of volatile compounds were studied in Emir grape must. Monocultures of W. saturnus and S. cerevisiae were also used. Inoculum level influenced the yeast growth, chemical composition and volatile compounds. W. saturnus began to die off with an increase in ethanol levels. The amounts of ethyl acetate and isoamyl acetate increased, but concentrations of 3-methylbutanol and ethanol decreased with increasing inoculum level from 5x106 to 1x108 cells/mL of W. saturnus in mixed cultures. Mono- and mixed culture fermentations inoculated with W. saturnus and S. cerevisiae yeasts did not form undesirable levels of flavour compounds. According to chemical composition and volatile compounds the differences between obtained wines were found significant. The addition of 5x106 cells/mL of W. saturnus appeared to a suitable inoculum level. It could be said that W. saturnus yeast can be used in mixed starter cultures with S. cerevisiae.

PRODUCTION OF PROBIOTIC YOGHURT (BIFILACT) BY USING SOME SPECIES OF LACTIC ACID BACTERIA

Probiotic lactic acid bacteria was used in manufacturing yoghurt from fortified whole milk. The product was stored under refrigeration for one week. The results showed that yoghurt manufacturing by using single or mixed strain bacteria was acceptable concerning acidity percent as it was 0.52 , 0.92 and 1.20 % using L.acidophilus , B.lactis and mixed culture , respectively for the fresh yoghurt. Total solid percentages of the fresh yoghurt were 16.47 , 16.48 and 15.93% for the three mentioned culture , respectively. Counts of Bifidobacteria in single strain starter and in mixed starter were higher followed by L.acidophilus in mixed starter, while counts were lowest in L.acidophilusin in single strain starter one week after storage counting were 81× 107 , 81× 107 , 36× 107 and 11× 107 c.f.u/ml. respectively, and these counts were within the acceptable ones of WHO and FAO values. Sensory evaluation showed that the product was acceptable for at least one week storage without any microbial or sensory changes which indicated that it can be used as a therapeutic product. Physical properties of the freeze dryed product were also studied and the results showed the ability for reconstruction and using it in capsule formulation.

Screening and selection of Bacillus spp. for fermented corticate soybean meal production

To screen and select the Bacillus spp. from Tua-nao of northern Thailand for fermented corticate soybean meal (FCSBM) production. After isolation of Bacillus spp. from Tua-nao was carried out, cellulase, hemicellulases (i.e., β-mannanase and xylanase) and phytase production by isolated Bacillus spp. were determined. B.subtilis isolate MR10 showed the highest β-mannanase, xylanase and phytase production at 280, 41 and 16Ug(-1) substrate, respectively, while the highest cellulase production was found in TK8 at 25Ug(-1) substrate. FCSBMs produced by single starter and mixed starter of both isolates showed the better properties than those of corticate soybean meal (CSBM), i.e., higher in soluble sugar, protein and phosphate content, smaller sugar molecules and better digestibility and absorbability than those of CSBM. Moreover, FCSBMs had no toxicity effect on mouse fibroblast cell line (3T3) but had an inhibitory effect on lung cancer cell line (CorL23). B.subtilis isolate MR10 and TK8 were selected for FCSBMs production because of their role as nutritional enhancer for CSBM and their safety. The results of this study were useful for FCSBM production process that can be applied as feed ingredient for monogastric animals.

Production of fermented goat beverage using a mixed starter culture of lactic acid bacteria and yeasts

Cow milk and goat milk can be used to produce fermented milk drinks, which have different properties. To use goat milk as a raw material, the fermentation conditions have to be carefully adapted to receive an optimal product. The aim of this study was to find the most appropriate starter culture for producing a health‐promoting goat beverage. The improvement of the quality of fermented goat beverage was done by selection of specific microbial strains and to define a ratio between the compounds of the starter culture. Goat milk was inoculated with starter cultures each containing two specified bacterial strains (Lacococcus lactis and L. lactis subsp. cremoris or Sterptococcus thermophilus and Lactobacillus delbruckii subsp. bulgaricus) as well as yeasts (Saccharomyces fragilis) in three concentrations 0.2, 0.4 or 0.6 g dm−3. The bacteria and yeasts growth and their metabolic profile was done by monitoring of the impedance. The fermentation process was characterized by pH value, alcohol content, color, and texture changes. Sensory evaluation of the final product was also performed. It was found that the use of L. lactis, L. lactis subsp. cremoris, and S. fragilis at 0.2 or 0.4 g dm−3 gave the most satisfying results and can be successfully utilized in goat milk fermentation.

Detection and identification of wild yeast in Koumiss

Koumiss is a slightly alcoholic fermented mare’s milk beverage, originally obtained by using a natural mixed starter of lactic acid bacteria and yeasts. Yeast is an important component of Koumiss processing which can affect the aroma, texture, as well as the nutrients beneficial to human health, but few reports have examined the yeast ecology of local ecosystems. The purpose of this study was to isolate and identify the yeast present in Koumiss from three representative regions of China using a polyphasic method. A total of 655 yeast isolates were obtained from 96 Koumiss samples collected from three regions in China. Koumiss harbored yeast populations at 5–7 log CFU/ml. Twelve different yeast species belonging to nine genera were detected in the Koumiss samples tested, including Candida pararugosa, Dekkera anomala, Geotrichum sp., Issatchenkia orientalis, Kazachstania unispora, Kluyveromyces marxianus, Pichia deserticola, Pichia fermentans, Pichia manshurica, Pichia membranaefaciens, Saccharomyces cerevisiae and Torulaspora delbrueckii. Kluyveromyces marxianus, Kazachstania unispora and Saccharomyces cerevisiae were the dominant species present in this traditional fermented dairy product. This study is the first to identify the yeast communities associated with Koumiss in China. The results enrich our knowledge of yeast in Koumiss, give us a more complete picture of the microbial diversity in Koumiss and can be used to promote the development of the local dairy industry.

Increased Qualities of Doenjang (Korean soybean paste) Prepared with Mixed Starter Cultures and Its Inhibitory Effects on Induction of Acute Colitis in Mice

We prepared Doenjang with single starter (Aspergillus oryzae) and mixed starter cultures (Aspergillus oryzae, Bacillus subtilis‐SKm and Lactococcuslactis‐GAm.), and compared their qualities and functionalities. The amino type nitrogen contents and the enzyme activities such as protease, lipase and α‐amylase increased in Doenjang when the mixed starters used. The DPPH free radical scavenging and antiproliferative activities in HT‐29 human colon cancer cells were also greater in Doenjang using the mixed starters than the other. Then, its preventive effects on acute colitis were investigated using in vivo colitis model. Colitis was induced by administration of 4% dextran sulfate sodium (DSS) to drinking water for 7 days in BALB/C mice. Doenjang were orally administered at 2 g/kg dose daily until sacrifice. The loss of body weight and shortening of colon length improved in the animals receiving Doenjang using the mixed starters as compared with the colitis‐induced control. Administration of the Doenjang also ameliorated histological score of the colon in DSS‐induced colitis mice. In addition, the treatment of Doenjang significantly repressed the proinflammatory cytokines of TNF‐α, IL‐12, IL‐17α and INF‐γ and the expression of inflammation related genes, such as COX‐2 and iNOS in comparison with the colitis‐induced control (P<0.05). This research is supported by the National Institute of Agricultural Science and Technology, Rural Development Administration, Korea.

Interaction between Lactococcus lactis and Lactococcus raffinolactis during growth in milk: Development of a new starter culture

Many milk fermentations use mixed cultures of lactic acid bacteria. To select a new mixed starter culture, 100 acid-producing bacterial strains were isolated from raw cow milk. Of these, 13 strains identified as belonging to the genera Lactococcus, Lactobacillus, Leuconostoc, or Weissella (based on phenotypic and genotypic tests) were assessed for a symbiotic effect between pairs of isolated strains during growth in milk. Among the strains tested, a mixed culture of Lactococcus lactis ssp. lactis strain 54 and Lactococcus raffinolactis strain 37 stimulated greater acid production during fermentation than occurred with pure fermentation. This stimulatory effect was not observed in milk supplemented with yeast extract or glucose or in constituted medium. Addition of a cell-free filtrate from milk fermented by strain 54 increased acid production by strain 37; however, the converse effect was not observed. The increased acid production by this mixed culture was, therefore, due to stimulation of strain 37 by metabolic products of strain 54, suggesting that the interaction between strains 54 and 37 is commensal. Analysis with a taste-sensing system indicated that fermented milk containing the mixed culture was more acidic, had more anionic bitterness, had greater aftertastes of anionic bitterness and astringency, and was less salty and umami than milk containing the individual cultures. This study identifies a new commensal relationship between 2 lactococcal strains that are commonly used for making dairy products.

Influence of fermentation conditions of Brassica oleracea L. var. capitata on the volatile glucosinolate hydrolysis compounds of sauerkrauts

The influence of fermentation conditions on the volatile glucosinolate (GLS) hydrolysis products in two different white cabbage cultivars (Bronco and Megaton) was studied. Natural and induced fermentation using Lactobacillus plantarum, Leuconostoc mesenteroides or a mixed starter culture of both microorganisms were performed. Cabbage cv. Bronco was fermented at a concentration of 0.5% and 1.5% NaCl while cv. Megaton was fermented only at 0.5% NaCl. Four commercial sauerkrauts were also analysed in order to compare with the experimental products. No volatile GLS hydrolysis products were detected in raw cabbages. Fermentation caused the appearance of iberin (IB), iberin nitrile (IBN), allyl cyanide (AC), allyl isothiocyanate (AITC) and sulforaphane (SFN) in experimental sauerkrauts, while only IB, IBN and SFN were detected in the commercial ones. Megaton sauerkrauts presented higher volatile GLS derivative content than those from cv. Bronco. The content of these compounds was affected by the starter culture and the salt concentration and it was in the range of those reported as having beneficial effect. Hence, sauerkraut can be considered as a health-promoting food and its intake is highly advised for disease prevention.

Persistence of wild Streptococcus thermophilus strains on wooden vat and during the manufacture of a traditional Caciocavallo type cheese

The present work was undertaken to evaluate the influence of the wooden dairy plant equipment on the microbiological characteristics of curd to be transformed into Caciocavallo Palermitano cheese. Traditional raw milk productions were performed concomitantly with standard cheese making trials carried out in stainless steel vat inoculated with a commercial starter. Milk from two different farms (A and B) was separately processed. The wooden vat was found to be a reservoir of lactic acid bacteria (LAB), while unwanted (spoilage and/or pathogenic) microorganisms were not hosted or were present at very low levels. All microbial groups were numerically different in bulk milks, showing higher levels for the farm B. LAB, especially thermophilic cocci, dominated the whole cheese making process of all productions. Undesired microorganisms decreased in number or disappeared during transformation, particularly after curd stretching. LAB were isolated from the wooden vat surface and from all dairy samples, subjected to phenotypic and genetic characterization and identification. Streptococcus thermophilus was the species found at the highest concentration in all samples analyzed and it also dominated the microbial community of the wooden vat. Fourteen other LAB species belonging to six genera (Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Streptococcus and Weissella) were also detected. All S. thermophilus isolates were genetically differentiated and a consortium of four strains persisted during the whole traditional production process. As confirmed by pH and the total acidity after the acidification step, indigenous S. thermophilus strains acted as a mixed starter culture.

Microbial dynamics and biodiversity in table olive fermentation: culture-dependent and -independent approaches

The microbial ecology of the table olive fermentation process is a complex set of dynamics in which the roles of the lactic acid bacteria (LAB) and yeast populations are closely related, and this synergism is of fundamental importance to obtain high quality products. Several studies on the ecology of table olives, both in spontaneous fermentations and in inoculated ones, have focused on the identification and characterization of yeasts, as they play a key role in the definition of the final organoleptic profiles through the production of volatile compounds. Moreover, these are able to promote the growth of LAB, which is responsible for the stabilization of the final product through the acidification activity and the inhibition of the growth of pathogenic bacteria. The current empirical production process of table olives could be improved through the development of mixed starter cultures. These can only be developed after a deep study of the population dynamics of yeasts and LAB by means of molecular methods. Until now, most studies have exploited culture-dependent approaches to define the natural microbiota of brine and olives. These approaches have identified two main species of LAB, namely Lactobacillus plantarum and L. pentosus, while, as far as yeasts are concerned, the most frequently isolated genera are Candida, Pichia, and Saccharomyces. However, there are a few studies in literature in which a culture-independent approach has been employed. This review summarizes the state of the art of the microbial ecology of table olive fermentations and it focuses on the different approaches and molecular methods that have been applied.

Inhibitory effects of meju prepared with mixed starter cultures on azoxymethane and dextran sulfate sodium-induced colon carcinogenesis in mice.

Backgrounds:Meju is the main ingredient and the starter culture of traditional Korean fermented soybean foods; these fermented soybean products are well-known for their various health benefits, including anticancer effects. We developed the grain-type meju using probiotic mixed starter cultures to improve the qualities and functionalities of fermented soybean products, as well as the meju itself. In this study, the inhibitory effects of the grain-type meju were investigated in azoxymethane (AOM) and dextran sulfate sodium (DSS)-induced colon carcinogenesis mice model.Materials and Methods:AOM and DSS colon carcinogenesis was induced in female C57BL/6 mice and meju was orally administered for 4 weeks. The body weight, colon length, and colon weight of mice were determined, and colonic tissues were histologically observed. The serum levels of proinflammatory cytokines and the levels of inflammation- and apoptosis-related genes in colonic tissue were also analyzed.Results:The administration of meju using probiotic mixed starter cultures ameliorated the symptoms of colon cancer and reduced number of neoplasia, and reduced serum proinflammatory cytokine levels and iNOS and COX-2 expression levels in colonic tissue. It increased Bax and reduced Bcl-2 expression levels and increased p21 and p53 expression in colonic tissues.Conclusion:The meju showed inhibitory effects on the progression of colon cancer induced by AOM and DSS by ameliorating the symptoms of colon cancer, reducing the number of neoplasias and regulating proinflammatory cytokine levels and the expressions of inflammation- and apoptosis-related genes in the colonic tissue.

발효제 비율에 따른 현미식초의 품질특성

Brown rice vinegar was made from brown rice mash with different brewing starter addition rates in static culture, and its quality characteristics were investigated. As a result, the amounts of alcohol produced in the fermentation process were shown to be 9.1, 8.8, 8.6 and 8.5% in the Nuruk 75 : crude enzyme 25 (B), Nuruk 50 : crude enzyme 50 (C), Nuruk 25 : crude enzyme 75 (D) and Nuruk 0 : crude enzyme 100 (E), respectively. The higher the percentage of crude enzyme added was, the lower the alcohol content that was produced. Nuruk 100 : crude enzyme 0 (A) was appeared to contain the lowest alcohol content (7.7%). In addition, the titratable acidity in all the groups was about 0.7%. The final titratable acidity (BV) of brown rice vinegar made with static culture was the highest (approximately 5.2%). The initial pH appeared to be between 3.6~4.0 and steadily decreased as the fermentation progressed, and the pH was almost unchanged after 15 day fermentation. The examination of the changes in the organic acids showed that the acetic acid content was similar in all the groups, and that the single starter added (AV, EV) group had much more other organic acids than the mixed starters added (BV, CV, DV) group. From these results, the mixed starters (Nuruk and crude enzyme) added group appeared to be superior to the single starter added in terms of alcohol production ability and vinegar quality. As the future aging process, however, is expected to change the flavor components and sensory characteristics, studies on various quality factors of vinegar are needed.

A TWO‐STEP CONTROLLED LACTIC FERMENTATION OF CABBAGE FOR IMPROVED CHEMICAL AND MICROBIOLOGICAL QUALITIES*

ABSTRACTTraditionally, lactic fermentation of vegetables is spontaneous and uncontrolled leading to unstable and inconsistent product. The present work used Leuconostoc mesenteroides and Lactobacillus plantarum in a two‐step controlled fermentation to ensure shelf stable, fermented cabbage with 0.5% added salt concentration. The pH, titratable acidity, lactic bacterial counts, sugar utilization and amino acid profile of the final product were periodically monitored. The pH in controlled cabbage fermentation dropped to less than 4.5, within 60 h of fermentation and glucose was completely utilized in 15 days while about 15–18% of glucose remained in spontaneously fermented cabbage. The spontaneously fermented cabbage samples had a significantly lower content of amino acids such as tyrosine, histidine, threonine, alanine, valine, phenylalanine, isoleucine, leucine, methionine and developed off‐flavors at the end of 90 days of refrigerated storage. In contrast, controlled lactic fermentation stabilized the product in the first 15 days of fermentation and was organoleptically acceptable up to 3 months.PRACTICAL APPLICATIONSUse of defined single or mixed starters and precise control of the cabbage fermentation process go a long way in ensuring a consistent product without the need of any chemical preservatives or harsh processing conditions. The present work made use of the benefits of heterofermentative pathway to create a product similar to naturally fermented cabbage in terms of flavor and used the homofermentative pathway to stabilize the product and showed that it is possible to obtain a satisfactory and stable product by the two‐step controlled fermentation as far as commercializing and mechanizing the product is concerned. The amino acid contents are also shown to be more in controlled than spontaneously fermented cabbage.

APLIKASI STARTER KULTUR MURNI PADA PEMBUATAN DURIAN FERMENTASI

This study looks into the effect of pure culture starters isolated from tempoyak on fermented durian (Durio zibethinus) and its acceptability. The use of 5% pure culture starters and mixed cultures of Pediococcus acidilactici, Lactobacillus plantarum, Lactobacillus curvatus Leuconostoc mesentroides and Staphylococcus saprophyticus were analyzed in terms of acceptabilty by sensory panelists. Statistical analysis shows a significant difference among fermented durian with pure culture and mixed culture starters for attributes of texture and viscosity, aroma and flavor, acidity and general acceptability.

Effect of Select Parameters of the Sourdough Rye Fermentation on the Activity of Some Mixed Starter Cultures

The effect of select parameters (i.e., rye flour ash content, temperature, and dough yield) of the sourdough fermentation on the fermentation activity of different starter cultures (lactic acid bacteria Lactococcus lactis ssp. Lactis, Weissella confusa, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus helveticus, and yeast Kluyveromyces marxianus subsp. Marxianus) was determined. The major metabolic end products of fermentation (D, L-lactic acid, acetic acid, ethanol and glycerol) and the evolution of total phenolic content and folic acid during bread making were measured. Lactobacillus helveticus and Kluyveromyces marxianus allowed obtaining sourdoughs with the highest lactic acid/acetic acid ratios. The mixed starter culture with Lactococcus lactis and Saccharomyces cerevisiae generated the most important quantities of D/L lactic acid. The maximum values of ethanol concentration were obtained in case of the sourdoughs from whole rye flour fermented at lower temperature (30°C) with mixed starter cultures containing Sacchomyces cerevisiae. The fermentation process and type of starter culture are also tools to increase the bioactive compounds, enabling the increase of the phenolic content of the sourdough.

Monitoring a mixed starter of Hanseniaspora vineae–Saccharomyces cerevisiae in natural must: Impact on 2-phenylethyl acetate production

The effect of simultaneous or sequential inoculation of Hanseniaspora vineae CECT 1471 and Saccharomyces cerevisiae T73 in non-sterile must on 2-phenylethyl acetate production has been examined. In both treatments tested, no significant differences in Saccharomyces yeast growth were found, whereas non-Saccharomyces yeast growth was significantly different during all days of fermentation. Independently of the type of inoculation, S. cerevisiae was the predominant species from day 3 till the end of the fermentation. The dynamics of indigenous and inoculated yeast populations showed H. vineae to be the predominant non-Saccharomyces species at the beginning of fermentation in sequentially inoculated wines, whereas the simultaneous inoculation of S. cerevisiae did not permit any non-Saccharomyces species to become predominant. Differences found in non-Saccharomyces yeast growth in both fermentations influenced the analytical profiles of final wines and specifically 2-phenylethyl acetate concentration which was two-fold increased in sequentially inoculated wines in comparison to those co-inoculated. In conclusion we have shown that H. vineae inoculated as part of a sequential mixed starter is able to compete with native yeasts present in non-sterile must and modify the wine aroma profile.

Killer phenotype of indigenous yeasts isolated from Argentinian wine cellars and their potential starter cultures for winemaking

Of 31 yeasts, from different surfaces of two cellars from the northwest region of Argentina, 11 expressed killer activity against the sensitive strain Saccharomyces cerevisiae P351. Five of these killer yeasts were identified as S. cerevisiae by phenotypic tests and PCR-RFLP analysis. Two S. cerevisiae killer strains, Cf5 and Cf8, were selected based on their excellent kinetic and enological properties as potential autochthonous mixed starter cultures to be used during wine fermentation. They could dominate the natural microbiota in fermentation vats and keep the typical sensorial characteristics of the wine of this region.

Changes in transcription profiles reflect strain contributions to defined cultures of Lactococcus lactis subsp. cremoris during milk fermentation

Cheddar cheese production uses mixed starters composed of Lactococcus lactis subsp. cremoris strains with complementary or competing enzymatic activity. However, strain interactions within the same subspecies are difficult to investigate by conventional microbiological methods. This study uses fluorescent RNA arbitrarily primed PCR (FRAP-PCR) to analyze the association of three L. lactis subsp. cremoris strains (LL074, LL225, and LL390 with proteinase types: PI, PIII, and PI/PIII, respectively) by monitoring whole transcription profiles. The effect of strain association was demonstrated by distinguishing profiles obtained with pure cultures from those obtained with defined mixed cultures. Both strains LL225 and LL390 dominated culture activity when in dual culture with strain LL074. The three-strain starter was also dominated by LL225 and LL390, in an approximately equal ratio which was stable over 35 generations. Strain LL225 had a stronger inhibitory effect on the growth of strain LL074 than LL390 did, showing incompatibility between strains LL225 and LL074. A new economic and semi-quantitative single-nucleotide polymorphism detection technique was developed and validated the results obtained by FRAP-PCR. Strain disequilibrium detected by FRAP-PCR could be related to inhibition of strain LL074 with a PI-type proteinase by the LL225 strain with a PIII-type proteinase. Strain compatibility could be characterized using these methods, leading to an improved understanding of mixed culture association of lactococcal strains.

Microbial Ecology Studies of Spontaneous Fermentation: Starter Culture Selection for Prickly Pear Wine Production

A procedure for designing starter cultures for fermentation is illustrated for prickly pear wine production. The illustration includes kinetic studies on inoculated and spontaneous fermentation, microorganism identification studies based on molecular biology tools, and microbial ecology studies, which led to the selection of strains that are capable of synthesizing alcohol and desirable volatile compounds. Results show that a mixed starter inoculum containing Pichia fermentans and Saccharomyces cerevisiae leads to a fermented product that contains 8.37% alcohol (v/v). The gas chromatography and mass spectrometry (GC-MS) analysis shows the presence of 9 major volatile compounds (Isobutanol, Isopentanol, Ethyl acetate, Isoamyl acetate, Ethyl octanoate, Ethyl decanoate, Ethyl 9-decanoate, β-Phenylethyl acetate, and Phenylethyl alcohol) that have ethereal, fruity, aromatic notes that are considered to be essential for a fine wine flavor. These compounds harmonically synergize with the alcohol to produce a fermented product with a unique flavor and taste. Several assays using the mixed culture show that the process is stable, predictable, controllable, and reproducible. Moreover, the results show that a mixed culture leads to a broader range of aromatic products than that produced by a single, pure culture. Therefore, we conclude that combinations of Saccharomyces strains and non-Saccharomyces strains can be used to obtain high-quality fermented beverages from prickly pear juice.