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Effect of solvent fractions of kombucha tea on viability and invasiveness of cancer cells—Characterization of dimethyl 2-(2-hydroxy-2-methoxypropylidine) malonate …
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In the present study, cytotoxic and anti-invasive properties of solvent fractions of kombucha tea were studied. Kombucha tea was fractionated with chloroform, ethyl acetate and butanol. Three solvent extracts and the final aqueous phase were concentrated and used for anti-cancer study at 25, 50, 75 and 100 μg/mL concentrations. Results revealed that ethyl acetate fraction at a concentration of 100 μg/mL caused cytotoxic effect on 786-O and U2OS cells; reduced the activities of matrix metalloproteinase-2 (MMP-2) and-9 in 786-O cells and MMP-2 activity in A549 cells; and significantly reduced the cell invasion and cell motility in A549, U2OS and 786-O cells. Thus, ethyl acetate fraction was further purified by chromatographic studies and presence of dimethyl 2-(2-hydroxy-2-methoxypropylidene) malonate and vitexin were confirmed through IR, NMR and mass spectroscopic studies, which might be responsible for the observed anticancer property of kombucha tea.
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... Kombucha offers a wide range of health benefits, including its ability to regulate blood sugar levels (Aloulou et al., 2012;Chakravorty et al., 2019), combat microbial infections (Deghrigue et al., 2013;Villarreal-Soto et al., 2018), protect against oxidative stress (Choi et al., 2023;Fajrin et al., 2023), reduce inflammation (Villarreal-Soto et al., 2018), prevent neurodegenerative diseases, lower high blood pressure, aid in detoxification, protect the liver and kidneys, alleviate stress, lower cholesterol levels (Júnior et al., 2022), and potentially inhibit cancer growth (Jayabalan et al., 2011;Srihari et al., 2013;Kim et al., 2016;Abou-Taleb et al., 2017;Bul Bul et al., 2018;Salafzoon et al., 2018;Villarreal-Soto et al., 2019;Kaewkod et al., 2019, Kaewkod et al., 2022Ghandehari et al., 2021Ghandehari et al., 2022Rasouli et al., 2021Laureys et al., 2020de Miranda et al., 2022). Therefore, health practitioners should advocate for the consumption of Kombucha to treat and prevent diseases, particularly for cancer patients, due to its beneficial properties. ...
... Multiple studies have demonstrated the efficacy of Kombucha as an anticancer agent due to its various organic acids and flavonoid groups (De Filippis et al., 2018;Jayabalan et al., 2011). Nevertheless, the underlying molecular mechanisms have not been fully elucidated. ...
... This multifaceted approach of TCM in combating cancer underscores its importance as a complementary or alternative therapeutic option in contemporary oncology (Lao et al., 2014;Tang et al., 2022). Jayabalan et al. (2011) studied the effect of the solvent fraction of Kombucha tea on the viability and invasiveness of cancer cells. The research identified dimethyl 2-(2-hydroxy-2-methoxypropylidine) malonate and vitexin as crucial components in this process. ...
... The fermentation process in the making of kombucha can be considered natural due to the addition of a certain amount of a previous kombucha fermentation which can be called back-slopping. Back slopping is a technique that is also utilised in the production of sourdough, fermented meats and cereals, water kefir, and milk kefir, among other natural food fermentation processes (Rasu Jayabalan et al., 2011). The fermentation process for kombucha normally takes 8 to 14 days at room temperature, during which time the characteristics of the sweetened tea evolve significantly. ...
... Tea polyphenols also have been proven to be the most impact towards anticancers by inhibiting the propagation of cancer cells (Rasu Jayabalan et al., 2011, 2014. The other beneficial chemical component such as vitamin C, glucuronic acid, gluconic acid, and lactic acid also helps in diminishing the incidence of stomach cancer (Xia et al., 2019). ...
Kombucha tea is a type of tea that undergoes fermentation through the inclusion of tea leaves, sugar, and a symbiotic culture of bacteria and yeast (SCOBY). The fermentation process typically spans a period of around 10 to 15 days. The utilization of this substance dates back to ancient times, mostly for medicinal purposes. The review has demonstrated that kombucha tea has distinctive characteristics, including anticancer, antioxidant, antimicrobial, and antifungal activities. The chemical component of kombucha plays an important role in kombucha tea which enhances the pharmacological activities such as acetic acid, lactic acid, phenolics and flavonoids. This paper presents an analysis of recent findings pertaining to the potential health advantages of kombucha, along with an examination of the chemical constituents and metabolites generated during the fermentation process. Additionally, further recommendations are provided for future investigations in this field.
... Similarly, the addition of Komagataeibacter rhaeticus (NAIMCC TB-3976) and yeast, specifically Brettanomyces bruxellensis (CAP 9), increased the acceptability of the roselle blended kombucha. However, Jayabalan et al. (39) revealed that kombucha tea ethyl acetate fractions contained two cytotoxic and anti-invasive chemicals, dimethyl 2-(2-hydroxyl -2 methoxypropyllidene malonate and vitexin. While experimenting on the prolonged fermentation studies of kombucha by Chen and Liu (40) using black tea as a fermentative substrate, organic acids such as gluconic acid were produced after 6th day of fermentation and the concentration of acetic acid increased very slowly, deciding the fruity taste of the final product. ...
Kombucha is a fermented drink with a range of medicinal benefits prepared from sweetened tea infusion (Camellia sinensis), which is cultured symbiotically with yeast and acetic acid bacteria. In the present investigation, kombucha was prepared from sugared black tea extract blended with aqueous calyx extract of Roselle (Hibiscus sabdariffa) @15% and fermented with cultures viz., Komagataeibacter rhaeticus (NAIMCCTB-3976) and Brettanomyces bruxellensis (CAP9) at 35°C. The floating water insoluble mat of kombucha was observed under a scanning electron microscope revealing the cellulosic nanofibrils secreted by K. rhaeticus. The total phenolic and flavanoid content, DPPH and ABTS activity of roselle calyx blended kombucha was significantly higher than black tea kombucha. Further, the compounds present in kombucha when analyzed by Fourier Transform Infra-Red Spectroscopy denoted the presence of carbonyl compounds, aromatic olefinic compounds, ketones, aldehydes, and esters. The different bioactive metabolites formed during fermentation were elucidated using Gas Chromatography-Mass Spectrometry and the major compounds excited within the retention time of 45 min with maximum peak area were13-hexyloxacyclotridec-10-en-2-one (37.64%), palmitins such as 1,3 dipalmitin (6.42%), glycidyl palmitate (3.30%), organic acids such as undecanedioic acid, linoleic acid, acetic acid (3.88%) etc., The results proved that blending black tea extract with 15% roselle calyx extract as a substrate for kombucha fermentation was highly accepted with an organoleptic score of 95% with improved functional properties than black tea extract kombucha alone.
... ABTS free radical-scavenging ability (%) = [1 − (As − Ab)/Ac] × 100 (2) where As is sample +ABTS reagent, Ab is sample +ddH 2 O, Ac is ddH 2 O +ABTS reagent, and different concentrations of ascorbic acid were additionally prepared as the control group. ...
Kombucha, a functional beverage rich in glucuronic acid, is fermented in the presence of acetic acid bacteria and yeast as the primary microorganisms. Glucuronic acid is recognized for its various physiological benefits, such as detoxification, antioxidation, and anti-inflammation. To optimize the glucuronic acid content in kombucha, various strain combinations by selecting fermented sources were accomplished. According to the experimental results, kombucha produced through co-fermentation with Pichia anomala and Komagataeibacter hansenii, with glucose-added black tea as the carbon source, exhibited the highest glucuronic acid production. A response surface methodology found that under optimized conditions of a 12.27% (w/v) carbon source concentration, a 10.07% (w/v) substrate concentration, and a 28.4 °C temperature, the highest glucuronic acid production reached 80.16 g/L, which represented a 2.39-fold increase compared to the original kombucha. Furthermore, the total polyphenol content increased by 3.87-fold, while DPPH and ABTS free radical–scavenging capacities increased by 1.86- and 2.22-fold, respectively. To sum up, these observations reveal the potential for commercial production of glucuronic acid–enriched kombucha and contribute to the development of functional food products related to kombucha in the future.
... Indicators such as aspartate and alanine transaminase, alkaline phosphatase in plasma, and the quantity of malondialdehyde in the plasma and liver tissues of albino rats in the experiment were used as supporting evidence. 17 Pioneering efforts to identify important volatiles/ metabolites by Jayabalan et al. 18 found that the ethyl acetate fractions of black tea kombucha contained two chemicals, dimethyl 2-(2-hydroxy-2-methoxypropylidene) malonate and vitexin, which have the ability to kill cells and prevent their invasion. These chemicals have shown anti-cancer effects in human lung carcinoma, osteosarcoma, and renal carcinoma cell lines. ...
Microbial cellulose, especially the bacterial cellulose produced by symbiotic co-cultures of acetic acid bacteria and yeast (SCOBY) that exists in a mutualistic interaction opens plausible strategies in the field of food as well as sustainable regenerative eco-system and waste management. Cultivated on sweetened black tea, the mutually proliferating bacteria (Acetobacter xylinum, A. xylinoides, and Bacterium gluconicum) and yeast strains (Schizosaccharomyces pombe, Saccharomycodes ludwigii and Saccharomyces cerevisiae) produces a fermented liquor along with the floating bacterial cellulosic pellicle called as Kombucha. This review explores the possible applications of kombucha SCOBY to use bacterial cellulose-based engineered living materials, commercial superabsorbent spheres by various marketing ventures like food, pharmaceutics, biomedical applications for bio-sensing and bio-catalysis, crop biostimulants, biocontrol agents in the management of plant and animal illnesses, post-harvest management in crops, water purification, pollutant detection, environmental biotechnology, and production of SCOBY from alternative substrates and agrarian waste management. The plausible use of bacterial cellulose hydrogels in dryland agriculture for their exceptional water-absorbing capability, eco-friendly nature, capacity to break down naturally, and compatibility with other living organisms is also elaborated in this paper. Furthermore, diverse microbial species to enhance the variety and functional properties of SCOBY, health benefits and its influence on human welfare is vividly discussed in the paper. The very in-depth study on the uses of SCOBY also paves way for the research exploration of this under-utilized microbial boon in food and farm sector for circular based regenerative agriculture in near future.
This study was conducted to develop kombucha with better functionality. The developed kombucha (CK) was prepared using the sugar extracts from fruits of Cudrania tricuspidata (Carrière) Bureau ex Lavallée instead of the sugar, which is used as a substrate for SCOBY in conventional kombucha (K). During fermentation, the soluble solids content significantly decreased in CK compared to K, and the pH change decreased rapidly in CK compared to K. On the 14th day of fermentation, the weight of the SCOBY in CK was higher than that in K. Immediately after preparation, K contained only sucrose, but CK contained sucrose, glucose, and fructose. SCOBY appears to use glucose and fructose preferentially during fermentation. K contained acetic acid and citric acid right after preparation. However, as fermentation progressed, the composition changed to acetic acid, citric acid, and lactic acid. At the same time, CK initially consisted of citric acid, lactic acid, and acetic acid. However, acetic acid and citric acid increased but lactic acid decreased significantly on the 14th day of fermentation. In the cytotoxicity studies, the CK showed a proliferation-promoting effect on normal lung cells (MRC-5) and strong cytotoxicity against human lung cancer cells (A549). These results suggest that the kombucha made from sugar extracts of C. tricuspidata fruits can be used as a more functional beverage than regular kombucha.
This study was conducted to develop kombucha with better functionality. The developed kombucha (CK) was prepared using the sugar extracts from fruits of Cudrania tricuspidata (Carrière) Bureau ex Lavallée instead of the sugar, which is used as a substrate for SCOBY in conventional kombucha (K). During fermentation, the soluble solids content significantly decreased in CK compared to K, and the pH change decreased rapidly in CK compared to K. On the 14th day of fermentation, the weight of the SCOBY in CK was higher than that in K. Immediately after preparation, K contained only sucrose, but CK contained sucrose, glucose, and fructose. SCOBY appears to use glucose and fructose preferentially during fermentation. K contained acetic acid and citric acid right after preparation. However, as fermentation progressed, the composition changed to acetic acid, citric acid, and lactic acid. At the same time, CK initially consisted of citric acid, lactic acid, and acetic acid. However, acetic acid and citric acid increased but lactic acid decreased significantly on the 14th day of fermentation. In the cytotoxicity studies, the CK showed a proliferation-promoting effect on normal lung cells (MRC-5) and strong cytotoxicity against human lung cancer cells (A549). These results suggest that the kombucha made from sugar extracts of C. tricuspidata fruits can be used as a more functional beverage than regular kombucha.
Kombucha is obtained by fermentation of tea and sugar by bacteria and yeast. This beverage has several beneficial effects, including antimicrobial activity. The objective of this study was to evaluate the antifungal activity of Kombucha extracts against dermatomycosis-causing fungi. Different types of Kombucha (black, white, green tea) and times of fermentation (10, 20, 30 days) were evaluated. Kombuchas were filtered, and extracts obtained by liquid-liquid extraction with ethyl acetate. Minimum inhibitory concentrations (MIC) were determined against clinical isolates of dermatophytes Trichophyton rubrum and T. mentagrophytes by broth microdilution. Toxicity tests and high-performance liquid chromatography (HPLC) were also performed. MIC ranged from 62.5–2000 μg/mL. Green tea Kombucha and 10-days fermentation extracts showed the best results. Toxicity was observed only for white tea Kombucha. Despite the chromatographic profiles, further studies are needed to characterize the active molecules. However, Kombucha extracts may constitute a viable alternative for new treatment strategies against dermatomycosis.
The effects of tea polyphenols on the invasion of highly metastatic human fibrosarcoma HT1080 cells through a monolayer of human umbilical vein endothelial cells (HUVECs) and the accompanying basal membrane were investigated. Among the tea polyphenols tested, epicatechin gallate (ECg), epigallocatechin gallate (EGCg), and theaflavin strongly suppressed the invasion of HT1080 cells into the monolayer of HUVECs/gelatin membrane, whereas epicatechin, epigallocatechin, tea flavonols, tea flavones, and gallate derivatives had no effect. Both theaflavin-digallate and theasinensin D showed a weak invasion inhibitory effect. ECg significantly inhibited the invasion without cytotoxicity against cancer cells and HUVECs. Ester-type catechins (ECg and EGCg) and theaflavin strongly suppressed the gelatin degradation mediated by matrix metalloproteinase (MMP) 2 and MMP-9, which were secreted into the conditioned medium of HT1080 cells. In conclusion, among the tea polyphenols tested, ECg was considered to be the agent with the most potential antimetastasis activity because it inhibited invasion in the absence of cytotoxicity. Keywords: Invasion; tea polyphenols; epicatechin gallate; human fibrosarcoma HT1080; matrix metalloproteinases
Kombucha tea is a fermented tea beverage produced by fermenting sugared black tea with tea fungus (kombucha). Tea polyphenols which includes (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin (EGC), (-)-epigallocatechin gallate (EGCG) and theaflavin (TF) have been reported to possess various biological activities. The present study focused on changes in content of organic acid and tea polyphenols in kombucha tea prepared from green tea (GTK), black tea (BTK) and tea manufacture waste (TWK) during fermentation. Concentration of acetic acid has reached maximum up to 9.5 g/l in GTK on 15th day and glucuronic acid concentration was reached maximum upto 2.3 g/l in BTK on 12th day of fermentation. Very less concentration of lactic acid was observed during the fermentation period and citric acid was detected only on 3rd day of fermentation in GTK and BTK but not in TWK. When compared to BTK and TWK very less degradation of EGCG (18%) and ECG (23%) was observed in GTK. TF and thearubigen (TR) were relatively stable when compared to epicatechin isomers. The biodegradation of tea catechins, TF and TR during kombucha fermentation might be due to some unknown enzymes excreted by yeasts and bacteria in kombucha culture.
Kombucha tea is a fermented tea beverage produced by fermenting sugared black tea with tea fungus (kombucha). Free-radical scavenging abilities of kombucha tea prepared from green tea (GTK), black tea (BTK) and tea waste material (TWK) along with pH, phenolic compounds and reducing power were investigated during fermentation period. Phenolic compounds, scavenging activity on DPPH radical, superoxide radical (xanthine-xanthine oxidase system) and inhibitory activity against hydroxyl radical mediated linoleic acid oxidation (ammonium thiocyanate assay) were increased during fermentation period, whereas pH, reducing power, hydroxyl radical scavenging ability (ascorbic acid-iron EDTA) and anti-lipid peroxidation ability (thiobarbituric assay) were decreased. From the present study, it is obvious that there might be some chances of structural modification of components in tea due to enzymes liberated by bacteria and yeast during kombucha fermentation which results in better scavenging performance on nitrogen and superoxide radicals, and poor scavenging performance on hydroxyl radicals.
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The biological activities of theaflavin (TF), theaflavin gallate (TFG) and theaflavin digallate (TFdiG) from black tea and (–)-epigallocatechin 3-gallate (EGCG) and (–)-epigallocatechin (EGC) from green tea were investigated using SV40-immortalized (33BES) and Ha- ras gene transformed (21BES) human bronchial epithelial cell lines. Growth inhibition and cell viability were measured by trypan blue dye exclusion assay following 24 h treatment with the tea polyphenols. TFdiG, EGC and EGCG displayed comparable inhibitory effects on the growth of 21BES cells, with estimated IC 50 values of 22–24 μM. TFG exhibited a lower inhibitory activity (IC 50 37 μM) and TF was even less effective (IC 50 47 μM) in this cell line. A similar effect was also observed in 33BES cells. These results suggest that the gallate structure of theaflavins is important for growth inhibition. Exposure of 21BES cells to 25 μM TFdiG, EGC and EGCG for 24 h led to induction of cell apoptosis/death as determined by the Annexin V apoptosis assay. With TFdiG treatment cell death occurred early, and quickly peaked at 8–12 h. Morphological observations showed that TFdiG-treated cells appeared irregular in shape, with cytoplasmic granules, suggesting a cytotoxic effect. On the other hand, EGC and EGCG showed a lag phase before a rapid increase in apoptosis between 16 and 24 h, without any marked morphological changes, which was similar to that induced by H 2 O 2 . TFdiG, EGC and EGCG induced similar amounts of H 2 O 2 formation in 21BES cells. Exogenously added catalase significantly prevented EGC- and EGCG-induced cell apoptosis, but did not prevent TFdiG-induced cell death, suggesting that H 2 O 2 is involved in the apoptosis induced by EGCG and EGC, but not in TFdiG-induced cell death. EGCG and TFdiG were shown to decrease c-jun protein phosphorylation in 21BES cells. Such inhibition is expected to result in lowered AP-1 activity, which may contribute to the growth inhibitory activity of tea polyphenols.
Previous studies in our laboratory have shown that the major green tea polyphenol, (–)-epigallocatechin-3-gallate (EGCG), suppressed autophosphorylation of epidermal growth factor (EGF) receptor induced by EGF in human A431 epidermoid carcinoma cells. In this study, we examined the inhibitory effects of black tea polyphenols, including theaflavin (TF-1), a mixture (TF-2) of theaflavin-3-gallate (TF-2a) and theaflavin-3′-gallate (TF-2b), theaflavin-3,3′-digallate (TF-3) and the thearubigin fraction on the autophosphorylation of the EGF and PDGF receptors in A431 cells and mouse NIH3T3 fibroblast cells, respectively. First, we examined the effects of these polyphenols on the proliferation of A431 and NIH3T3 cells. Both EGCG and TF-3 strongly inhibited the proliferation of A431 and NIH3T3 cells more than the other theaflavins did. In cultured cells with pre-treatment of tea polyphenol, TF-3 was stronger than EGCG on the reduction of EGF receptor and PDGF receptor autophosphorylation induced by EGF and PDGF, respectively. Other theaflavins slightly reduced the autophosphorylation of the EGF and PDGF receptors; furthermore, TF-3 could reduce autophosphorylation of the EGF receptor (or PDGF receptor) even with co-treatment with EGF (or PDGF) and TF-3, but EGCG was inactive under these conditions. In addition, TF-3 was stronger than EGCG in blocking EGF binding to its receptor. These results suggest that not only the green tea polyphenol, EGCG, but also the black tea polyphenol, TF-3, have an antiproliferative activity on tumor cells, and the molecular mechanisms of antiproliferation may block the growth factor binding to its receptor and thus suppress mitogenic signal transduction.
A tetrazolium salt has been used to develop a quantitative colorimetric assay for mammalian cell survival and proliferation. The assay detects living, but not dead cells and the signal generated is dependent on the degree of activation of the cells. This method can therefore be used to measure cytotoxicity, proliferation or activation. The results can be read on a multiwell scanning spectrophotometer (ELISA reader) and show a high degree of precision. No washing steps are used in the assay. The main advantages of the colorimetric assay are its rapidity and precision, and the lack of any radioisotope. We have used the assay to measure proliferative lymphokines, mitogen stimulations and complement-mediated lysis.