Dynamic changes in aromas and precursors of edible fungi juice: mixed lactic acid bacteria fermentation enhances flavor characteristics.

BACKGROUND: Lactic acid bacteria (LAB) fermentation technology has been increasingly used in the deep processing of edible fungi. However, the flavor profiles of edible fungi products after mixed LAB fermentation have received less attention and how aromas changes during the mixed LAB fermentation are still open questions. In the present study, fermented Hericium erinaceus and Tremella fuciformis compound juice (FHTJ) was prepared by mixed LAB strains. We aimed to systematically monitor the dynamic changes of aromas and precursors throughout the fermentation process and a data-driven association network analysis was used to tentatively illustrate the mechanisms of formation between aromas and their precursors. RESULTS: Mixed LAB fermentation could enrich the aroma profile of FHTJ, reducing the unpleasant flavors such as nonanal and 1-octen-3-ol, as well as increasing the floral flavors such as ethyl acetate and α-pinene. Partial least squares-discriminant analysis and relative odor activity values revealed that 11 volatile chemicals were recognized as aroma-active markers. Volcano plot analysis showed that 3-octen-2-one (green flavor) was the key aroma-active marker in each stage, which was down-regulated in fermentation stages I, II and IV, whereas it was up-regulated in stage III. 3-Octen-2-one was significantly negatively correlated with organic acids, particularly pyruvate (r2 = -0.89). Ethyl caprylate (floral flavor) was up-regulated in the late fermentation stage, and showed a negative correlation with sugar alcohols and a positive correlation with organic acids, especially tartaric acid (r2 = 0.96). CONCLUSION: The present study demonstrates the beneficial effect of mixed LAB fermentation on flavor characteristics, providing guidance for fermented edible fungi juice flavor quality monitoring and control. © 2024 Society of Chemical Industry.

Effects of Different Non-Saccharomyces Strains in Simultaneous and Sequential Co-Fermentations with Saccharomyces cerevisiae on the Quality Characteristics of Kiwi Wine.

With the increasing awareness of health, more people have shown a preference for low-alcohol beverages. Seeking various methods to improve the quality of kiwi wine is now a major research interest in the wine industry. In this study, kiwi wine was fermented by Saccharomyces cerevisiae and different non-Saccharomyces strains (Torulaspora delbrueckii, Kluyveromyces thermotolerans, Pichia fermentans) in three methods (pure fermentation, simultaneous, and sequential co-fermentation). The physicochemical characteristics, color parameters, phenolic profiles, total phenolic content (TPC), antioxidant activities, organic acids, and taste sense of the different wines were evaluated to determine the effects of different yeasts and fermentation methods on the quality of the kiwi wine. Results indicated that co-fermentation reduced the contents of alcohol while enhancing the lightness of the kiwi wine. The TPC of sequential co-fermentation with K. thermotolerans/S. cerevisiae was significantly higher than that of their simultaneous co-fermentation. Compared to K. thermotolerans/S. cerevisiae, the antioxidant activities were increased by co-fermentation of T. delbrueckii/S. cerevisiae and P. fermentans/S. cerevisiae. Principal component analysis showed that kiwi wines fermented by different yeasts and inoculation methods could be separated and grouped. Correlation analysis presented positive correlations of phenolic composition, antioxidant activities, and color intensity. This study provided theoretical guidance for co-fermentation of non-Saccharomyces/S. cerevisiae and accelerated the industrialization process of kiwi wine.

Cheese Whey Protein and Blueberry Juice Mixed Fermentation Enhance the Freeze-Resistance of Lactic Acid Bacteria in the Freeze-Drying Process.

The effects of MRS, whey protein and blueberry alone, and mixed fermentation on the survival rate of lactic acid bacteria under various freeze-drying conditions were investigated. The surface structure of the freeze-dried powders was also investigated to explore the anti-freezing protection mechanism of mixed whey protein and blueberry fermentation on the bacteria. It was found that the mixed fermentation medium of blueberry and whey protein has a protective effect on the freeze-drying bacteria and is better than the traditional MRS and whey protein medium. The optimal concentration of blueberry juice addition was 9%. The survival rate of the pre-freezing temperature at -80 °C was higher than at -20 °C after the pre-freezing and freeze-drying processes. The freeze-drying thickness of 0.3 cm could improve the survival rate of the bacteria. The Fourier transform infrared spectroscopy results indicated the interaction between the whey protein, anthocyanins, and the surface composition of the lactic acid bacteria.

Effects of cofermentation of Saccharomyces cerevisiae and different lactic acid bacteria on the organic acid content, soluble sugar content, biogenic amines, phenol content, antioxidant activity and aroma of prune wine.

To determine the effect of cofermentation of Saccharomyces cerevisiae and different LABs on prune wine quality, this study compared phenolic compounds, organic acids, soluble sugars, biogenic amines and volatile flavor compounds among different treatments. The results showed that inoculation of LAB increased DPPH and total flavonoid content. Malic acid content was reduced in HS, HB and HF. Histamine content in S, F and B was lower than the limits in French and Australian wines. 15 phenolic compounds were identified. Yangmeilin and chlorogenic acid were detected only in HS, HF and HB. 51 volatile flavor compounds were identified, esters being the most diverse and abundant. 14 volatile flavor compounds with OAV > 1 contributed highly to the aroma of prune wine. 9 chemical markers including resveratrol, rutin, and catechin were screened to explain intergroup differences by OPLS-DA. This study provides new insights into the processing and quality analysis of prunes.

Advancements and challenges for brewing aroma-enhancement fruit wines: Microbial metabolizing and brewing techniques.

Aroma, a principal determinant of consumer preference for fruit wines, has recently garnered much attention. Fruit wines brewing was concomitant with complex biochemical reactions, in which a variety of compounds jointly contribute to the aroma quality. To date, the mechanisms underlying the synthesis of aroma compounds and biological regulation methods in fruit wines have remained ambiguous, hindering the further improvement of fruit wines sensory profiles. This review provides a detailed account of the synthesis and regulatory mechanisms of typical aroma compounds and their contributions to the characteristics of wines. Additionally, Comprehensive involves between microflora and the formation of aroma compounds have been emphasized. The microflora-mediated aroma compounds evolution can be controlled by key fermentation techniques to protect and enhance. Meanwhile, the genes impacting key aroma compounds can be identified, which provide references for the rapid screening of aroma-enhanced strains as well as target formation of aroma by modifying relative genes.

Evaluation of the effect of Torulaspora delbrueckii on important volatile compounds in navel orange original brandy using E-nose combined with HS-SPME-GC-MS.

Simultaneous inoculation of non-Saccharomyces cerevisiae during the alcoholic fermentation process has been found to be an effective strategy for enhancing wine flavor. This study aimed to investigate the effect of Torulaspora delbrueckii NCUF305.2 on the flavor of navel orange original brandy (NOOB) using E-nose combined with HS-SPME-GC-MS. The results showed a significant increase (p < 0.05) in the sensitivity of NOOB to W5C, W3C, W1S, and W3S sensors by mixed fermentation (MF). Esters in NOOB increased by 4.13%, while higher alcohols increased by 21.93% (p < 0.001), terpenes and others increased by 52.07% and 40.99% (p < 0.01), respectively. Notably, several important volatile compounds with relative odor activity values above 10 showed an increase. Sensory analysis revealed that a more pronounced citrus-like flavor and higher overall appearance scores were found in MF than in pure fermentation (PF). These findings offer valuable theoretical guidance for enhancing the quality of fruit brandies.

Improving aromatic higher alcohol acetates in wines by co-fermentation of Pichia kluyveri and Saccharomyces cerevisiae: growth interaction and amino acid competition.

BACKGROUND: Higher alcohol acetates (HAAs) are potent aroma-active esters that impart desirable fruity and floral aromas. However, the conversion of higher alcohol precursors into HAAs is extremely low in winemaking. To investigate the underlying yeast-yeast interaction on targeted improvement of aromatic HAAs, we evaluated fermentation activity, cell viability, amino acid consumption and HAA production when Pichia kluyveri and Saccharomyces cerevisiae were inoculated concurrently or sequentially. RESULTS: Pichia kluyveri PK-21 possessed the ability to survive and increased HAA level up to 5.2-fold in mixed fermentation. Such an increment may benefit from the efficient conversion of higher alcohol precursors into HAAs (>27-fold higher than S. cerevisiae). During mixed fermentation, the two yeasts exhibited crucial interactions regarding cell growth and amino acid competition. Saccharomyces cerevisiae dominated over the co-inoculated P. kluyveri by efficient uptake of amino acids and biomass production. However, this dominance decreased in sequential fermentation, where P. kluyveri growth increased due to the consumption of preferred amino acids prior to S. cerevisiae. Pearson correlation analysis indicated that phenylalanine and aspartic acid may act as positive amino acids in boosting P. kluyveri growth and HAA production. Laboratory-scale winemaking validated the fermentation performance of P. kluyveri in sequential inoculum, resulting in a balanced aroma profile with enhanced floral and tropical fruity characteristics in the final wines. CONCLUSION: This study proposes a microbial, non-genetically engineered approach for targeted increase of HAA production in winemaking and the findings provide new insights into yeast-yeast interactions. © 2024 Society of Chemical Industry.

Effects of Torulaspora delbrueckii and Saccharomyces cerevisiae Co-Fermentation on the Physicochemical and Flavor Compounds of Huaniu Apple Cider.

The effects of different fermentation methods utilizing Torulaspora delbrueckii 1004 and Saccharomyces cerevisiae 32169 on the physicochemical properties, organic acid content, polyphenol and flavonoid concentrations, antioxidant activity, and volatile aroma compounds of Huaniu apple cider were investigated in this study. Employing methods of single inoculation, co-inoculation, and sequential inoculation, it was found that sequential fermentation exhibited strong fermentative power in the initial stages, effectively reducing the content of soluble solids and achieving a balanced composition of malic, succinic, and citric acids while maintaining a lower titratable acidity. Sequential inoculation was observed to significantly enhance the total polyphenols and flavonoids, as well as the antioxidant capacity (p < 0.05). Specifically, in the synthesis of volatile aroma compounds, sequential inoculation significantly enhanced the richness and diversity of the Huaniu apple cider's aromas, particularly in terms of the concentration of ester compounds (p < 0.05). Principal component analysis further confirmed the superiority of sequential inoculation in terms of aroma component diversity and richness. The findings of this study suggest that sequential inoculation of fermentation with non-Saccharomyces and S. cerevisiae is an effective strategy for optimizing the flavor characteristics of Huaniu apple cider, offering valuable theoretical support and practical guidance for enhancing cider quality and fostering the development of new products.

Effects of Non-Saccharomyces Yeasts and Their Pairwise Combinations in Co-Fermentation with Saccharomyces cerevisiae on the Quality of Chunjian Citrus Wine.

Non-Saccharomyces (NSc) yeasts have great potential in improving wine qualities. In this study, two NSc and two Saccharomyces cerevisiae (Sc) samples were tested on their performance of mono-inoculated and composite culture in the fermentation of Chunjian citrus wine. The cell count, Brix degree, total sugar, total acidity, alcohol level, pH value, color intensity (CI), and tonality were determined to evaluate the contribution of NSc to the quality of citrus wine in the mixed fermentation. Volatile compounds were analyzed by HS-SPME-GC-MS, and sensory evaluation was carried out. During the 9-day fermentation, the mixed-culture wine exhibited a higher cell concentration than the pure culture. After the fermentation, mixed-culture wine specifically decreased the concentrations of unfavorable volatile compounds, such as isobutanol and octanoic acid, and increased favorable volatile compounds, including ethyl octanoate, ethyl decanoate, and phenylethyl acetate. The quality category of the citrus wine was improved compared with the Sc mono-inoculated wines, mainly in regard to aroma, retention, and sweetness. The study shows that the mixed fermentation of NSc and Sc has positive impacts on reducing alcohol level and total acidity and increasing CI. The present work demonstrates that the mixed fermentation of NSc and Sc has enormous beneficial impacts on improving the quality of citrus wine.

Staphylococcus inoculation enhances the sensorial attributes of Chinese bacon by coordinating the composition of flavor compounds through amino acid metabolism.

In this study, we aimed to uncover the potential underlying mechanisms of the flavor modulation of Chinese bacon by Staphylococcus. To that end, taste-enhancing S. cohnii WX-M8 and S. saprophyticus MY-A10 screened from Chinese bacon were used to investigate the effects of their individual and mixed fermentations and their synergistic fermentation with Lactobacillus plantarum BL-1 on the sensorial attributes, physicochemical properties, microbial diversity, and volatile compounds (VOCs) of Chinese bacon. Our results revealed that S. cohnii WX-M8 and S. saprophyticus MY-A10 significantly increased a* (redness) and Aw and reduced thiobarbituric acid reactive substances (TBARS) when fermented in a mixture. Moreover, they promoted the formation of esters, aldehydes (especially straight-chain aldehydes), and phenolic compounds through pathways related to amino acid metabolism, enhancing sensorial attributes. While synergistic fermentation with L. plantarum BL-1 resulted in an improved a* (redness) of Chinese bacon, and the increased microbial metabolism of the carbohydrate and lipid metabolic pathways, the increase in TBARS and the higher content of acidic volatiles, led to a change in the composition of the flavor substances. The advantage of co-fermentation of Staphylococci in sensory attributes can be attributed to their capability to metabolize amino acids and associates. These findings provide insights into the role of Staphylococcus as a starter in regulating bacon flavor.

Effect of mixed fermentation of Lactiplantibacillus plantarum and Lactiplantibacillus pentosus on phytochemical and flavor characteristics of Wallace melon juice.

BACKGROUND: Melons (Cucumis melo L.) are among the most commonly consumed fruits but they are highly susceptible to mechanical damage and rot during storage and transportation. New processed products are needed to avoid postharvest fruit loss and to increase health benefits. Fermentation is an effective means of utilizing the nutrients and improving flavor. RESULTS: Fermented melon juice (MJ) was prepared using three potential probiotics Lactiplantibacillus plantarum CICC21824 (LP), Lactiplantibacillus plantarum GB3-2 (LG), and Lactiplantibacillus pentosus XZ-34 (LX). The nutrition, flavor characteristics, and digestive properties of different fermented MJs were compared. The results demonstrated that, in comparison with mono-fermentation, mixed fermentation by LG and LX could increase the level of organic acids and phenolic acids. Correspondingly, antioxidant capacity was improved significantly and positively correlated with p-coumaric acid and cinnamic acid content. The production of alcohols and acids was more strongly enhanced by mixed culture fermentation, whereas mono-fermentation reduced the content of esters, especially ethyl acetate and isopropyl acetate. Aldehydes and ketones increased significantly in fermented MJ, and damascenone and heptanal could be the characteristic aroma compounds. CONCLUSION: Mixed fermented MJ provides more beneficial phytochemicals, better flavor, and stronger antioxidant properties than mono-fermentation. © 2024 Society of Chemical Industry.

The effect of mixed culture fermentation of Saccharomyces cerevisiae and Saccharomyces cerevisiae var. diastaticus on fermentation parameters and flavor profile.

Belgian Saisons and Lambics are two well-known examples in the brewing industry of mixed fermentations, combination of two or more yeast and/or bacteria strains. The purpose of this study was to determine the impact different pitch rates of Saccharomyces cerevisiae (traditional brewing yeast) and S. cerevisiae var. diastaticus (a variant associated with Belgian styles) had on the fermentation kinetics and concentration of the volatile compounds in the finished beers. A series of brews were performed utilizing ratios of S. cerevisiae and diastaticus. The fermentations were heavily monitored, and a model was used to fit fermentation variables. It was found that mixed fermentations produced behaviors that were predictable and proportional to the mixture ratios. As expected, the pure cultural fermentations of diastaticus had a slower fermentation midpoint (M) at 45.45 h versus 28.28 h for S. cerevisiae with the mixed ones falling in between the two. Flavor and aroma play a key role in the acceptability of beer. The mixed fermentations showed a combination of the two different yeast strains aromatic profiles. When combined, there was a strong linearity between alcohols (R2  = 0.94), esters (R2  = 0.89), and the overall total (R2  = 0.91) volatile compounds. PRACTICAL APPLICATION: Modeling is a widely utilized tool in several different fields. The purpose of this research is to apply modeling techniques to describe the fermentation speed and flavor profile of a mixed fermentation between S. cerevisiae and diastaticus. The equations from this data can be used by brewers for product development purposes to make beers with certain flavor profiles within a desired timeframe.

Wine aroma modification by Hanseniaspora uvarum: A multiple-step strategy for screening potential mixed starters.

Hanseniaspora uvarum is a prevalent yeast species in vineyards. However, its application in grape wine fermentation remains limited. This study used culture-dependent and -independent approaches to investigate the dynamics of H. uvarum during the spontaneous fermentation of Cabernet Sauvignon grapes. The results revealed that H. uvarum constituted 77.49 % of the non-Saccharomyces yeast population during fermentation. An indigenous strain, QTX-C10, was isolated from the 148 H. uvarum strains using a multistep screening strategy. The 1:1 co-inoculation of QTX-C10 with Saccharomyces cerevisiae proved to be an optimal strategy for mixed fermentation, resulting in a 48.54 %-59.55 % increase in ethyl esters in Cabernet Sauvignon wine and a 96.94 %-110.92 % increase in Chardonnay wine. Furthermore, this approach reduced the acetic acid levels by 12.50 %-17.07 % for Cabernet Sauvignon wine and 10.81 %-17.78 % for Chardonnay wine. Additionally, increased ethyl ester content may enhance the tropical fruit flavor of Cabernet Sauvignon wines.

Unveiling the Aromas and Sensory Evaluation of Hakko Sobacha: A New Functional Non-Dairy Probiotic Fermented Drink.

At the dawn of a food transition encouraging the consumption of healthy and sustainable non-dairy probiotic products, the development of a fermented functional drink based on Sobacha is considered. Sobacha is an infusion of roasted buckwheat seeds widely consumed in Asian countries for its health benefits. As fermentation improves the nutritional and organoleptic status of grains, the mixed fermentation process involved in the development of kombucha beverages (fermented sweet tea) is conducted by inoculating a symbiotic culture of bacteria and yeasts into the transposable matrix (Sobacha instead of tea). Sobacha, a healthy pseudo-cereal matrix with promising aromas, could be fermented to potentially develop an innovative drink, named "Hakko Sobacha". This neologism would reveal the fermented character of the infusion, Hakko meaning fermented in Japanese. Considering the beverage characterization, the kinetics of the volatile organic compound syntheses were determined using stir-bar sorptive extraction followed by gas chromatography coupled to mass spectrometry analysis. Odor-active compounds were theoretically calculated to estimate the flavor composition. Finally, sensory analyses highlighted the appreciation and preferences of the consumer towards the beverages. The fermentative yield differences observed between the two buckwheat concentration modalities tested seemed to be correlated with the sugar and nutrient levels available from the starch (buckwheat) matrix. Having characterized Hakko Sobacha, this study proposed the possibility of developing new beverages by monitoring the fermentative process. This should enable improved control and enhancement of their sensorial properties, which could in turn lead to greater customer acceptability.

Isolation of Aroma-Producing Wickerhamomyces anomalus Yeast and Analysis of Its Typical Flavoring Metabolites.

In this study, 21 strains of aroma-producing yeast were isolated from Sichuan paocai juice of farmers in western, eastern and southern Sichuan. One strain, Y3, with the best aroma-producing characteristics, was screened using an olfactory method and a total ester titration method, and was identified as Wickerhamomyces anomalus. The total ester content of Y3 fermentation broth was as high as 1.22 g/L, and there was no white colonies or film on the surface. Meanwhile, the Y3 strain could tolerate 14% salt concentration conditions and grow well in a pH range of 3-4. Through sensory analysis, the fermented mustard with a ratio of Lactiplantibacillus plantarum to Y3 of 1:1 showed the highest overall acceptability. Ethyl acetate with its fruit and wine flavor was also detected in the fermented Sichuan paocai juice with a mixed bacteria ratio of 1:1, analyzed with SPME-GC-MS technology, as well as phenylethyl alcohol, isobutyl alcohol, isothiocyanate eaters, myrcene and dimethyl disulfide. These contributed greatly to the unique flavor of Sichuan paocai. In general, Wickerhamomyces anomalus Y3 enhanced the aroma of the fermented Sichuan paocai.

Dynamical changes of tea metabolites fermented by Aspergillus cristatus, Aspergillus neoniger and mixed fungi: A temporal clustering strategy for untargeted metabolomics.

Dark tea fermentation involves various fungi, but studies focusing on the mixed fermentation in tea remain limited. This study investigated the influences of single and mixed fermentation on the dynamical alterations of tea metabolites. The differential metabolites between unfermented and fermented teas were determined using untargeted metabolomics. Dynamical changes in metabolites were explored by temporal clustering analysis. Results indicated that Aspergillus cristatus (AC) at 15 days, Aspergillus neoniger (AN) at 15 days, and mixed fungi (MF) at 15 days had respectively 68, 128 and 135 differential metabolites, compared with unfermentation (UF) at 15 days. Most of metabolites in the AN or MF group showed a down-regulated trend in cluster 1 and 2, whereas most of metabolites in the AC group showed an up-regulated trend in cluster 3 to 6. The three key metabolic pathways mainly composed of flavonoids and lipids included flavone and flavonol biosynthesis, glycerophospholipid metabolism and flavonoid biosynthesis. Based on the dynamical changes and metabolic pathways of the differential metabolites, AN showed a predominant status in MF compared with AC. Together, this study will advance the understanding of dynamic changes in tea fermentation and provide valuable insights into the processing and quality control of dark tea.

Mixed fermentation and electrospray drying for the development of a novel stabilized wheat germ powder containing highly viable probiotic cultures.

Nondairy fermented probiotic powder was developed based on stabilized wheat germ through mixed fermentation (Lactobacillus acidophilus and Lactobacillus plantarum) and electrospraying process. In the first step, the effect of mixed fermentation on lipase and lipoxygenase activity of wheat germ was investigated. The results showed a significant reduction in the activity of both enzymes (82.72% for lipase and 72% for lipoxygenase), therefore, mixed fermentation effectively stabilizes the wheat germ. In the next step, after the preparation of the solutions for drying process and investigating the physical properties (surface tension, electrical conductivity, and viscosity) of the solutions, the electrosprayability of the samples was evaluated at different conditions and revealed that 18 kV applying voltage, 0.3 flow rate, and 12 cm distance between tip to collector was the best for electrospraying the 20% solution of fermented wheat germ with morphologically most semi-uniform particles. Finally, the viability of the probiotics after drying process and during the storage at 25°C was examined. The number of initial cells counted as 14.48 ± 0.2 log cfu/g and the viability studies showed 0.55 log cfu/g decrease in the number of viable bacteria from initial count as a result of the electrospraying process. Furthermore, 7.86 ± 0.03 log cfu/g in freeze-dried and 9.05 ± 0.45 log cfu/g in electrosprayed samples survived after 70 days of storage.

Adjustment of impact phenolic compounds, antioxidant activity and aroma profile in Cabernet Sauvignon wine by mixed fermentation of Pichia kudriavzevii and Saccharomyces cerevisiae.

Mixed fermentation using saccharomyces cerevisiae and non-saccharomyces cerevisiae has become one of the main research strategies to improve wine aroma. Hence, this study applied the mixed fermentation technique using Pichia kudriavzevii and Saccharomyces cerevisiae to brew Cabernet Sauvignon wine and to investigate the effects of inoculation timing and inoculation ratio on the polyphenolics, antioxidant activity and aroma of the resulting wine. The results showed that mixed fermentation significantly improved the amounts of flavan-3-ols. In particular, S1:5 had the highest amounts of (-)-catechin and procyanidin B1 (73.23 mg/L and 46.59 mg/L), while S1:10 had the highest (-)-epicatechin content (57.95 mg/L). Meanwhile, S1:10 showed the strongest FRAP, CUPRAC and ABTS + activities (31.46 %, 25.38 % and 13.87 % higher than that of CK, respectively). In addition, mixed fermentation also increased the amounts of phenylethanol, isoamyl alcohol and ethyl esters, which enhanced the rose-like and fruity flavor of wine. This work used a friendly non-saccharomyces cerevisiae alongside appropriate inoculation strategies to provide an alternative approach for improved wine aroma and phenolic profile.

Characterization of Aroma Active Compound Production during Kombucha Fermentation: Towards the Control of Sensory Profiles.

Since the sensorial profile is the cornerstone for the development of kombucha as a beverage with mass market appeal, advanced analytical tools are needed to gain a better understanding of the kinetics of aromatic compounds during the fermentation process to control the sensory profiles of the drink. The kinetics of volatile organic compounds (VOCs) was determined using stir bar sorptive extraction-gas chromatography-mass spectrometry, and odor-active compounds were considered to estimate consumer perception. A total of 87 VOCs were detected in kombucha during the fermentation stages. The synthesis of mainly phenethyl alcohol and isoamyl alcohol probably by Saccharomyces genus led to ester formation. Moreover, the terpene synthesis occurring at the beginning of fermentation (Δ-3-carene, α-phellandrene, γ-terpinene, m- and p-cymene) could be related to yeast activity as well. Principal component analysis identified classes that allowed the major variability explanation, which are carboxylic acids, alcohols, and terpenes. The aromatic analysis accounted for 17 aroma-active compounds. These changes in the evolution of VOCs led to flavor variations: from citrus-floral-sweet notes (geraniol and linalool domination), and fermentation brought intense citrus-herbal-lavender-bergamot notes (α-farnesene). Finally, sweet-floral-bready-honey notes dominated the kombucha flavor (2-phenylethanol). As this study allowed to estimate kombucha sensory profiles, an insight for the development of new drinks by controlling the fermentation process was suggested. Such a methodology should allow a better control and optimization of their sensory profile, which could in turn lead to greater consumer acceptance.

Headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) and odor activity value (OAV) to reveal the flavor characteristics of ripened Pu-erh tea by co-fermentation.

Introduction: Pu-erh tea is a geographical indication product of China. The characteristic flavor compounds produced during the fermentation of ripened Pu-erh tea have an important impact on its quality. Methods: Headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) and odor activity value (OAV) is used for flavor analysis. Results: A total of 135 volatile compounds were annotated, of which the highest content was alcohols (54.26%), followed by esters (16.73%), and methoxybenzenes (12.69%). Alcohols in ripened Pu-erh tea mainly contribute flower and fruit sweet flavors, while methoxybenzenes mainly contribute musty and stale flavors. The ripened Pu-erh tea fermented by Saccharomyces: Rhizopus: Aspergillus niger mixed in the ratio of 1:1:1 presented the remarkable flavor characteristics of flower and fruit sweet flavor, and having better coordination with musty and stale flavor. Discussion: This study demonstrated the content changes of ripened Pu-erh tea's flavor compounds in the fermentation process, and revealed the optimal fermentation time. This will be helpful to further understand the formation mechanism of the characteristic flavor of ripened Pu-erh tea and guide the optimization of the fermentation process of ripened Pu-erh tea.