Fermentation Period Research Articles

Evaluation of the fermentative activity of Saccharomyces cerevisiae HG1.3 and condition for threeleaf cayratiawine preservation

Threeleaf cayratia(Cayratia trifolia L.) is a wild climbing vine species that has potential raw material for wine production. Berries are green and turn purple when ripe, widely distributed in the Mekong delta provinces of Vietnam. This study aimed to investigate the threeleaf cayratiawine fermentation activity of Saccharomyces cerevisiae HG1.3 at the volumes of 1, 2, and 4 litres, along with examining the colour stability of the wine under different storage temperatures and packaging methods. The results showed that the fermentation volumes of 2 and 4 litres contained 12.37 and 12.45% (v/v) of ethanol contents, respectively. At the storage conditions of cool temperature (2-4°C)in the dark bottle, threeleaf cayratiawine has a stable colour and good sensory evaluation for colour and taste clarity after 4 weeks, with the optical transmittance value at 550 nm is 0.419%. In addition, threeleaf cayratiawine was clarified by the addition of 0.5% pectinase for 6 days after the fermentation period, then filtered and supplemented with 0.2% citric acid. The final product meets the Vietnamese standards (TCVN 3217-79 and TCVN 7045:2002).

Optimization of antioxidant activity and characterization of probiotic cascara beverage fermented using water kefir grain [Optimasi aktivitas antioksidan dan karakterisasi minuman probiotik cascara hasil fermentasi menggunakan bibit kefir air

Cascara is a beverage derived from dried coffee cherry pulp, known to have good functional value but with a short shelf life. Fermentation is a technique that can enhance cascara beverages' shelf life and quality and their functional value due to the presence of probiotics and increased antioxidant compounds. The study aimed to optimize the fermentation conditions of cascara tea with water kefir seeds to improve the microbiota content of Lactobacillus sp, Bifidobacterium sp, and Saccharomyces cerevisiae and the antioxidant activity of the beverage. The method in this study used Response Surface Methodology (RSM) with a design selection of Central Composite Design (CCD), using two independent variables, namely fermentation time (X1) and Grain Concentration (X2), with a response of antioxidant activity (%). The findings indicated that a fermentation period of 48 hours and a grain concentration of 8.58% resulted in a probiotic drink exhibiting the maximum antioxidant activity of 65.53%. This optimum treatment resulted in a drink with IC50 antioxidant activity of 173.66 ppm, total phenol 477.24 mg GAE/mL, brightness value (L*) 30.69, redness (a*) 14.06, yellowness (b*) 29.57, pH 3.57, TPT 8.80 (ᵒBrix), viscosity 1.62 mPa.s, total LAB 8.54 log CFU/ml, and total yeast & mold 10.93 log CFU/ml.

Intermolecular interactions between malvidin-3-O-glucoside and caffeic acid: Structural and thermodynamic characterization and its effect on real wine color quality

The copigmentation effect between malvidin-3-O-glucoside and caffeic acid was comprehensive inquiry on the model wine solution, theoretical simulation and real wine. Thermodynamic parameters were determined by UV/Visible spectroscopy and Isothermal titration calorimetry (ITC). Theoretical data were obtained employing a dispersion-corrected density functional approach. The effects in real wines were investigated by adding the caffeic acid during different fermentation periods. Results shown that the copigmentation reaction between caffeic acid and malvidin-3-O-glucoside is a spontaneous exothermic reaction driven by hydrogen bonding and dispersions forces. Computations show that the polyhydroxyl sugar moiety and phenolic hydroxyl groups are the key active sites. The addition of caffeic acid in post-alcohol fermentation samples evidences an improving color characteristics in the wine.

A chromosome-scale genome provides new insights into the typical carotenoid biosynthesis in the important red yeast Rhodotorula glutinis QYH-2023 with anti-inflammatory effects

Rhodotorula spp. has been studied as one powerful source for a novel cell factory with fast growth and its high added-value biomolecules. However, its inadequate genome and genomic annotation have hindered its widespread use in cosmetics and food industries. Rhodotorula glutinis QYH-2023, was isolated from rice rhizosphere soil, and the highest quality of the genome of the strain was obtained at chromosome level (18 chromosomes) than ever before in red yeast in this study. Comparative genomics analysis revealed that there are more key gene copies of carotenoids biosynthesis in R. glutinis QYH-2023 than other species of Rhodotorula spp. Integrated transcriptome and metabolome analysis revealed that lipids and carotenoids biosynthesis was significantly enriched during fermentation. Subsequent investigation revealed that the over-expression of the strain three genes related to carotenoids biosynthesis in Komagataella phaffii significantly promoted the carotenoid production. Furthermore, in vitro tests initially confirmed that the longer the fermentation period, the synthesized metabolites controlled by R. glutinis QYH-2023 genome had the stronger anti-inflammatory properties. All of the findings revealed a high-quality reference genome which highlight the potential of R. glutinis strains to be employed as chassis cells for biosynthesizing carotenoids and other active chemicals.

Flavor Precursor and Volatile Compounds Formation of Unfermented Cocoa Beans Hydrolyzed by Papain

Fermentation is notably crucial stage in the production of cocoa flavor precursors. Nevertheless, some cocoa plantations in Indonesia skip fermentation as the raw material is relatively small, and the fermentation period is extremely long. This study aimed to improve the flavor precursors in unfermented cocoa beans (var. forastero). Unfermented cocoa beans were hydrolyzed using papain at different concentrations and incubation times. The conditions to obtain the highest degree of hydrolysis were 3.3 U/mL (papain conc.) at 10 h incubation. After papain treatment, it exhibited lower reducing sugars and polyphenol content than unfermented cocoa beans. Besides, hydrophobic amino acids such as phenylalanine, valine, leucine, and isoleucine was increased. After roasting, volatile compounds for chocolate aroma were also presented. However, pyrazines, aldehydes, and esters, were still less than those in fermented cocoa beans. The results proved that the papain hydrolysis of unfermented cocoa beans can improve their flavor precursors and volatile compounds.

Antifungal efficacy of natural antimicrobial substances during the fermentation and storage of hardaliye

Hardaliye is a non-alcoholic traditional fermented beverage produced using black grapes, mustard seeds, sour cherry leaves, and sodium benzoate. In this study, the effect of using plant materials (quince leaves, green tea, mulberry leaves, linden flowers, horseradish roots) as an alternative to sodium benzoate for inhibiting yeast growth to produce additive-free hardaliye was investigated. Hardaliye samples were produced using the selected test materials, microbiological and chemical analyses were performed during fermentation (20 °C for 18 days) and storage (4 °C for 2 months) periods. Additionally, the antifungal effects of test material extracts against Saccharomyces cerevisiae in grape juice and Malt Extract Broth (MEB), as well as the survival of S. cerevisiae in grape juice containing plant materials were examined. Only mustard seed extract had inhibitory and fungicidal effects at 25 mg/mL on S. cerevisiae in MEB among the tested plant material extracts and the effect increased when used in combination with grape juice. The total phenolic contents of the hardaliye samples were in the range of 2119.26–8653.33 mg gallic acid equivalents (GAE)/L, and antioxidant activities were in the range of 30.24–93.15%. The numbers of S. cerevisiae decreased below the detection limit (<1 log CFU/mL) after 1 day in grape juice containing horseradish (5%), mustard seeds (1.5%) or sodium benzoate (0.1%). This study indicated that crushed mustard seeds at increased concentrations (1.5%), mustard seeds water extract (7.5 mg/mL), or horseradish roots (5%) have the potential to be used in the production of hardaliye as an alternative to sodium benzoate.

Texture Profile Analysis of Lamtoro Gung (Leucaena leucocephala ssp. Glabrata (Rose) S. Zarate) Tempeh

Lamtoro Gung seeds are low in fat content but rich in protein. Lamtoro Gung is often used as an alternative protein source. Lamtoro Gung is usually taken as a prepared meal containing grains and nuts. It is germinated and utilized in a variety of dishes, including the production of tempeh. However, nothing has been done to explain the textural profile of Lamtoro Gung tempeh, which is made by fermentation processes in various packaging materials such as plastic, banana leaves, and teak leaves. This study aims to evaluate the textural characteristics of Lamtoro Gung tempeh produced using different packaging materials and fermentation periods. This study contributes to understanding how packaging variations affect the texture of Lamtoro Gung tempeh. Tempeh made from teak leaf packaging and fermented for four days offered better texture profile hardness (195.38 N), cohesiveness (0.46), gumminess (81.57 N), and chewiness (54.99 N) higher than banana leaves; hardness (49.90 N), cohesiveness (0.29), gumminess (14.52 N), and chewiness (10.14 N) but similar to plastic; hardness (165.28 N), cohesiveness (0.45), gumminess (74.23 N), and chewiness (50.35 N). Therefore, packaging material impacts the texture profile of tempeh fermented from Lamtoro Gung seeds. Sensory evaluation of the tempeh would need to be investigated to check consumer acceptance of the product.

Developing a High-Umami, Low-Salt Soy Sauce through Accelerated Moromi Fermentation with Corynebacterium and Lactiplantibacillus Strains.

The traditional fermentation process of soy sauce employs a hyperhaline model and has a long fermentation period. A hyperhaline model can improve fermentation speed, but easily leads to the contamination of miscellaneous bacteria and fermentation failure. In this study, after the conventional koji and moromi fermentation, the fermentation broth was pasteurized and diluted, and then inoculated with three selected microorganisms including Corynebacterium glutamicum, Corynebacterium ammoniagenes, and Lactiplantibacillus plantarum for secondary fermentation. During this ten-day fermentation, the pH, free amino acids, organic acids, nucleotide acids, fatty acids, and volatile compounds were analyzed. The fermentation group inoculated with C. glutamicum accumulated the high content of amino acid nitrogen of 0.92 g/100 mL and glutamic acid of 509.4 mg/100 mL. The C. ammoniagenes group and L. plantarum group were rich in nucleotide and organic acid, respectively. The fermentation group inoculated with three microorganisms exhibited the best sensory attributes, showing the potential to develop a suitable fermentation method. The brewing speed of the proposed process in this study was faster than that of the traditional method, and the umami substances could be significantly accumulated in this low-salt fermented model (7% w/v NaCl). This study provides a reference for the low-salt and rapid fermentation of seasoning.

Production of red beetroot juice by different methods: Kinetics of microbial growth, sugar consumption, and acid production

As a fermentation method, the utilisation of starter culture is a common practice in industrial manufacturing, although spontaneous methods have been employed since ancient times. The objective of this study was to investigate the effect of different production methods on red beetroot juice (RBJ). For this purpose, as a starter culture, the probiotic Lactibasillus paracasei (Lc. paracasei) was inoculated into the RBJ samples after pasteurization. Also, the growth of cells, acid production, and substrate utilisation were monitored throughout the fermentation process of RBJ under two different methods of fermentation.The samples produced by the addition of Lc. paracasei demonstrated a slightly lower decrease in pH values in comparison to the samples obtained by the spontaneous method. The concentration of lactic acid (LA) and acetic acid (AA) at the end of fermentation reveals that Lc. paracasei exhibits a greater capacity for both LA and AA generation compared to the spontaneous method. The ratios of LA and AA molar concentrations of RBJ were determined to be 1.7 and 3.6 for the samples produced by adding Lc. paracasei and the spontaneous method, respectively. The samples produced by adding Lc. paracasei exhibited a greater consumption of sucrose. Both fermentation methods provide LAB counts exceeding 8 log CFU/mL at the end of fermentation.Time demonstrated a significant correlation with LA and AA in the method by adding Lc. paracasei (r = 0.942 and 0.745), respectively (p < 0.01). In both methods, it was demonstrated that while sucrose content decreased during the fermentation period, fructose and glucose content remained constant (p < 0.05).

Exploring the Relationships Between Bacterial Community, Taste-Enhancing Peptides and Aroma in Thai Fermented Fish (Pla-ra).

As a traditional Thai condiment, Pla-ra is used to add flavor and richness to dishes. Nine treatment combinations of Pla-ra formulations created from 3 types of fish (Mor fish, Kradee fish, and Mor + Kradee fish) and 4 different carbohydrate sources (none, rice bran, roasted rice, and rice bran─roasted rice mixture) were studied through a 12 month fermentation period (1, 3, 5, 7, 8, 9, 10, 11, and 12 months). 16S rRNA Next Generation Sequencing (NGS) and LC-MS/MS techniques were used to analyze the microbial diversity and identify taste-enhancing peptides. Descriptive sensory analysis was performed on the extracts of the 108 Pla-ra samples mixed in a model broth. Koku perception and saltiness-enhancing attributes were clearly perceived and dominant in all samples, even though glutamyl peptides, including γ-Glu-Val-Gly, were found at subthreshold levels. The samples from mixed fish and Mor fish fermented with roasted ground rice and rice bran for 12 months had the most typical Pla-ra odors and tastes and had high taste-enhancing activities. NGS analysis revealed the presence of bacteria containing a large number of protease and aminopeptidase genes in the samples. Bacillus spp., Gallicola spp., and Proteiniclasticum spp. correlated well with the generation of glutamyl and arginyl peptides and typical odors in the samples. These results confirmed the typical sensory quality of Pla-ra depended on protein sources, carbohydrate sources, and bacteria communities. Further optimization of the microbial composition found could lead to the development of starter cultures to control and promote flavor development in fermented fish products.

Dynamics of peptides released from cow milk fermented by kefir microorganisms during fermentation and storage periods

This study investigates the changes in peptides released during the fermentation and storage of cow milk with a kefir culture using porous graphitized carbon-liquid chromatography-tandem mass spectrometry (PGC-MS/MS). A comprehensive inventory of 695 distinct milk peptides from 98 milk proteins was determined, with 523 characterized within 24 h of fermentation. The quantitative evaluation of released peptides throughout the kefir fermentation process unveiled noteworthy variations. A total of 50 discrete bioactive peptides with various biological functions were determined throughout the fermentation stages. After fermentation, we identified 26 bioactive peptides within the resulting beverage, maintaining their stability throughout storage. In addition, during the storage periods, three new bioactive peptides emerged. The composition of bioactive peptides underwent alterations during fermentation but remained relatively constant during storage. Our study sheds light on the proteolytic activity of potential enzymes released into the milk samples from kefir cultures, contributing to the beverage's evolution during storage.

Deciphering the core microbiota in open environment solid-state fermentation of Beijing rice vinegar and its correlation with environmental factors.

Rice vinegar is a popular cereal vinegar worldwide and is typically produced in an open environment, and the ecosystem of solid-state fermentation is complicated and robust. The present study aimed to reveal the shaping force of the establishment of the ecosystem of Beijing rice vinegar, the core function microbiota and their correlation with critical environmental factors. [Correction added after first online publication on 29 May 2024; the word "worldwide" has been removed from the first sentence under the section Background.] RESULTS: The experimental findings revealed the changes in environmental factors, major metabolites and microbial patterns during Beijing rice vinegar fermentation were obtained. The major metabolites accumulated at the middle and late acetic acid fermentation (AAF) periods. Principal coordinates and t-test analyses revealed the specific bacterial and fungal species at corresponding stages. Kosakonia, Methlobacterium, Sphingomonas, unidentified Rhizobiaceae, Pseudozyma and Saccharomycopsis dorminated during saccharification and alcohol fermentation and early AAF, whereas Lactococcus, Acetobacter, Rhodotorula and Kazachstania dominated the later AAF stages. Canonical correspondence analysis of environmental factors with core microbiota. Temperature and total acid were the most significant factors correlated with the SAF bacterial profile (Pediococcus, Weissella, Enterococcus and Kosakonia). Ethanol was the most significant factor between AAF1 and AAF3, and mainly affected Acetobacter and Lactobacillus. Conversely, ethanol was the most significant factor in the SAF, AAF1 and AAF3 fungi communities; typical microorganisms were Saccharomyces and Malassezia. Furthermore, the predicted phenotypes of bacteria and their response to environmental factors were evaluated. In conclusion, the present study has provided insights into the process regulation of spontaneous fermentation and distinguished the key driving forces in the microbiota of Beijing rice vinegar fermentation. © 2024 Society of Chemical Industry.

Optimization of production and evaluation of Microbial kojic Acid obtained from Sugarcane Molasses (SCM) by Aspergillus sp.

Kojic acid (KA) is an organic acid that is generated by various fungi, particularly by Aspergillus species, as a secondary metabolite. The current study is aimed to determine the optimal conditions for the production of kojic acid from various fungal strains grown on agro-industrial wastes. After testing six fungal strains for their suitability for kojic acid production, Aspergillus oryzae (AUMC.64) and Aspergillus tamari (AUMC.43) were found to be the highest producers of KA. Three different agro-industrial wastes were screened as a fermentation media and sugar cane molasses showed the highest productivity for (KA). Aspergillus oryzae (AUMC.64), and Aspergillus tamari (AUMC.43) achieved the maximal production of kojic acid (25.91, 18.95 ± 0.001 g. L-1respectively) from sugarcane molasses (SCM) under optimum conditions of growth (10% solution of sugarcane molasses, pH 4.0 and fermentation period of 10 days). Also, the antimicrobial activities of KA produced by A. oryzae AUMC64 and A. tamari AUMC43 against the selected test strains of microorganisms Staphylococcus aureus, Bacillus cereus, Escherichia coli, Salmonella typhimurium were recorded. The maximum growth inhibition zone (20–13.2 mm) was observed on the cultures of Escherichia coli. Meanwhile the antioxidant activities of KA produced by A. oryzae AUMC64 and A. tamari AUMC43 was 79.1 and 62.42%, respectively.

Casein hydrolysate in naturally-fermented buckwheat sourdough: Effects on fermented and physicochemical characteristics, texture, and bacterial microbial composition

The effect of a casein hydrolysate (CH) on the fermentation and quality of a naturally-fermented buckwheat sourdough (NFBS) were investigated, through assessing the fermentation characteristics, carbohydrate and protein degradation, texture, and bacterial composition of NFBS. According to the assaying data, CH might both increase the amount of lactic acid bacteria by 2.62 % and shorten the fermentation period by at least 3 h, subsequently leading to enhanced degradation of carbohydrate and protein, accompanied by a softer texture. More importantly, CH increased the relative abundance of lactobacillus in NFBS, making it the dominant bacterial genus and inhibited the growth of spoilage bacteria. In addition, Spearman correlation analysis indicated that the pH value, lactic and acetic acid contents, carbohydrates, protease activity, and these textural indices like hardness, elasticity, and adhesion had a positive/negative correlation with the bacterial composition of NFBS (Spearman correlation coefficient: −0.93-0.95). CH was thus regarded to be helpful to NFBS processing and production mainly by shortening its fermentation time, improving its fermentation performance, causing a finer texture and microstructure, and changing bacterial composition.

Effects of Brassinosteroid Hormone on Chemical Composition of Okra (Abelmoschus esculentus L.) under Salt Stress

Following the scientific management of organic manure (poultry) may contribute to reducing environmental pollution and human health, as the current study was conducted with the aim of recycling poultry manure accumulated in the field of poultry farming at the research station - Faculty of Agriculture - Karma site and the impact of varying fermentation durations on organic manure, specifically animal (poultry manure) fermented by different fermentation methods and its impact on the rest of the factors to produce highly decomposed organic manure free of pathogens. Several traditional fermentation methods (compost and water immersion) were followed. and modern (bioreactor), and for a fermentation period of 90 days with an estimate of most of the qualitative characteristics of the organic manure produced and within periods of 15 - 30 - 45 - 60 - 75 - 90 days. The results of the experiment of incubation of poultry manure showed that all the studied traits have changed with the progress of fermentation time and in the same direction, the results demonstrated that fermentation in the bioreactor unit has given the highest values of total nitrogen, total phosphorus and potassium 32.53, 16.72, 20.86, g.kg-1 for the elements mentioned sequentially, while the lowest values of pH 6.68 and C/N ratio 9.96 compared to other fermentation methods.

Effect of fermentation periods of poultry manure on its characteristics when fermented with different ways

Following the scientific management of organic manure (poultry) may contribute to reducing environmental pollution and human health, as the current study was conducted with the aim of recycling poultry manure accumulated in the field of poultry farming at the research station - Faculty of Agriculture - Karma site and the impact of varying fermentation durations on organic manure, specifically animal (poultry manure) fermented by different fermentation methods and its impact on the rest of the factors to produce highly decomposed organic manure free of pathogens. Several traditional fermentation methods (compost and water immersion) were followed. and modern (bioreactor), and for a fermentation period of 90 days with an estimate of most of the qualitative characteristics of the organic manure produced and within periods of 15 - 30 - 45 - 60 - 75 - 90 days. The results of the experiment of incubation of poultry manure showed that all the studied traits have changed with the progress of fermentation time and in the same direction, the results demonstrated that fermentation in the bioreactor unit has given the highest values of total nitrogen, total phosphorus and potassium 32.53, 16.72, 20.86, g.kg-1 for the elements mentioned sequentially, while the lowest values of pH 6.68 and C/N ratio 9.96 compared to other fermentation methods.

Optimization of CO fermentation by Clostridium carboxidivorans in batch reactors: Effects of the medium composition

ObjectivesThe objective of this study was to investigate the effects of medium composition on CO fermentation by Clostridium carboxidivorans. The focus was to reduce the medium cost preserving acceptable levels of solvent production. MethodsYeast extract (YE) concentration was set in the range of 0–3 g/L. Different reducing agents were investigated, including cysteine-HCl 0.6 g/L, pure cysteine 0.6 g/L, sodium sulphide (Na2S) 0.6 g/L, cysteine-sodium sulphide 0.6 g/L and cysteine-sodium sulphide 0.72 g/L. The concentration of the metal solution was decreased down to 25 % of the standard value. Fermentation tests were also carried out with and without tungsten or selenium. ResultsThe results demonstrated that under optimized conditions, namely yeast extract (YE) concentration set at 1 g/L, pure cysteine as the reducing agent and trace metal concentration reduced to 75 % of the standard value, reasonable solvent production was achieved in less than 150 h. Under these operating conditions, the production levels were found to be 1.39 g/L of ethanol and 0.27 g/L of butanol. Furthermore, the study revealed that selenium was not necessary for C. carboxidivorans fermentation, whereas the presence of tungsten played a crucial role in both cell growth and solvent production. ConclusionsThe optimization of the medium composition in CO fermentation by Clostridium carboxidivorans is crucial for cost-effective solvent production. Tuning the yeast extract (YE) concentration, using pure cysteine as the reducing agent and reducing trace metal concentration contribute to reasonable solvent production within a relatively short fermentation period. Tungsten is essential for cell growth and solvent production, while selenium is not required.

Effect of Pediococcus acidilactici bioaugmentation on the quality improvement of milkfish (Chanos chanos) Bekasam

Bekasam is a traditional Indonesian fermented food usually composed of freshwater fish, salt, steamed rice as a carbohydrate source, and garlic, prepared by spontaneous fermentation for approximately one week up to 1 month, which gives it a distinctive scent and flavor. Milkfish is commonly used as a main ingredient due to its popularity in certain areas and the many possible variations of Bekasam. This study investigated the effect of Pediococcus acidilactici bioaugmentation on the quality of Bekasam. During the 12 days of production, the spontaneous milkfish Bekasam (SMB) was compared to the milkfish Bekasam with the addition of P. acidilactici (PAMB). The starter significantly impacted the properties of Bekasam, giving it a slightly brown appearance as a results of Maillard reaction and increasing the protein content (P&lt;0.05), indicating proteolytic activity during fermentation which directly affects the flavor and texture of the end product. The considerably increased protein content was a result of the lengthy fermentation period. Additionally, the fermentation starter resulted in a lower total plate count and increased growth of lactic acid bacteria than the non-starter (P&lt;0.05) Bekasam. Furthermore, the total sugar concentration reduced as fermentation progressed, indicating that the starter and the length of the fermentation period play a role in enhancing Bekasam’s quality and health-related properties.

Time-Course Metabolomic Analysis: Production of Betaine Structural Analogs by Fungal Fermentation of Seaweed.

Betaine structural analogs are compounds characterized by the presence of positive and negative charges in a single molecule and have been reported to have physiological properties, such as anti-inflammatory activities. In this study, we performed a metabolomic analysis of metabolite composition changes during the fermentation of Neopyropia yezoensis, an edible red alga, with Aspergillus oryzae for 72 h. The results indicated that three specific betaine structural analogs (betaine, stachydrine, and carnitine) exhibited significant changes in production by the end of the 72 h fermentation period. Time-course analysis suggested that betaine was generated from the precursor choline at 12-24 h during the late stage of fungal growth, while stachydrine was generated from the precursor-related compound glutamic acid at 48-72 h during the sporulation stage. However, the contribution of the precursor lysine to the increased production of carnitine during the 12-72 h period was unclear. This study provides useful information on the efficient production of betaine structural analogs by the fungal fermentation of seaweed as well as various other food materials.

Metabolomic kinetics investigation of Camellia sinensis kombucha using mass spectrometry and bioinformatics approaches

Kombucha is created through the fermentation of Camellia sinensis tea leaves, along with sucrose, utilizing a symbiotic consortium of bacteria and yeast cultures. Nonetheless, there exists a dearth of comprehensive information regarding the spectrum of metabolites that constitute this beverage. To explore this intricate system, metabolomics was used to investigate fermentation kinetics of Kombucha. For that, an experimental framework was devised to assess the impact of varying sucrose concentrations and fermentation temperatures over a ten-day period of kombucha fermentation. Following fermentation, samples were analyzed using an LC-QTOF-MS system and a distinctive metabolomic profile was observed. Principal component analysis was used to discriminate between metabolite profiles. Moreover, the identified compounds were subjected to classification using the GNPS platform. The findings underscore notable differences in compound class concentrations attributable to distinct fermentation conditions. Furthermore, distinct metabolic pathways were identified, specially some related to the biotransformation of flavonoids. This comprehensive investigation offers valuable insights into the pivotal role of SCOBY in driving metabolite production and underscores the potential bioactivity harbored within Kombucha.