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A new kinetic study of the uncatalyzed bromate oscillator using dark tea without pre-treatment was carried out. The difference of dark tea oscillating system with and without manganese ion as catalyst was compared. The reaction was studied with different starting concentrations of the reagents (sulfuric acid and potassium bromate) and the substrate of dark tea, and the results indicate that the concentration of dark tea has a wide range between 1 with 30 g/L in dark tea-KBrO3-H2SO4 system, the lowest concentration of potassium bromate and sulfuric acid are 0.028 mol/L and 0.24 mol/L. The effects of the temperature were also investigated and the activation energy of oscillation was calculated to 88.78 kJ/mol. In addition, the possible mechanism of the non-catalyzed oscillating reaction was also discussed briefly. [ABSTRACT FROM AUTHOR]

To enhance our understanding of Aspergillus cristatus, an important functional microorganism, the characteristics of its mitochondrial genome were analyzed and compared with related species. The mitochondrial genome of A. cristatus was determined to be 77,649 bp in length, with 15 protein-coding regions. Notably, its length surpassed that of the other species, primarily attributable to the intron length. Gene order exhibited significant variations, with greater conservation observed in the genus Penicillium compared to Aspergillus. Phylogenetic tree analyses indicated that the genera Aspergillus and Penicillium are closely related but monophyletic. Furthermore, the phylogenetic tree constructed based on protein-coding genes effectively distinguished all strains with high branching confidence. This approach provides a robust reflection of the evolutionary relationship between A. cristatus and its related species, offering potential for the development of molecular markers suitable for Aspergillus and Penicillium.

Dark tea refers to a kind of post-fermented product, and its quality and price vary owing to the distinct altitudes at which it grows. In this study, a novel method based on high performance liquid chromatography with a diode-array detector (HPLC-DAD) and an evaporative light scattering detector (HPLC-ELSD) was proposed for the classification of dark teas from distinct altitudes in China. Through implementing a strategy fusing feature-level data to construct a combined dataset, the classification performance of dark teas from distinct altitudes in China was evaluated after preprocessing. The results suggested that, through the feature fusion strategy, the identification accuracy rate increased from

Dark tea, rich in nutricines including tea polyphenols and free amino acids, is a kind of post-fermented tea. The potential application of nutricines against oxidative damage and senescence, which drives animal health maintenance and disease prevention, has attracted considerable interest. In this study, the effect of dark tea and its effects on longevity and defense against oxidative stress was investigated in the Caenorhabditis elegans (C. elegans) model. Under normal conditions, dark tea extended the lifespan without significant impairment of propagation. It also improved the motility, alleviated the fat accumulation and apoptosis. Additionally, orally administered dark tea could significantly decrease the level of reactive oxygen species (ROS) and resulted in a superior lifespan in H2O2-induced oxidative stressed C. elegans. In antioxidant assays in vitro, dark tea was found to be rich in strong hydroxyl, DPPH and ABTS+ free radical scavenging capacity. Interestingly, mRNA sequence analyses further revealed that dark tea may catalyze intracellular relevant oxidative substrates and synthesize antioxidants through synthetic and metabolic pathways. These results suggest that dark tea is worth further exploration as a potential dietary supplement for the maintenance of animal health and the prevention of related diseases. [ABSTRACT FROM AUTHOR]

The tea plant is a vital strategic forest resource in China. Dark tea produced from its leaves is an indispensable health-promoting product in western China due to its unique lipid-lowering function. Eurotium cristatum is the dominant strain in Fuzhuan brick tea (a variety of Anhua dark tea) and could produce many functional components, including lovastatin, a lipid-lowering compound. In this study, the lovastatin yield of dark tea was improved by breeding Eurotium cristatum using the protoplast fusion method. The experiments were carried out by inducing a fusion between inactivated Eurotium cristatum JH1205 and Monascus CICC5031. Among the 92 fusants screened the HPLC method, four strains (A4, A36, A54, and A76) with higher lovastatin production (more than three times as high) were obtained. The A76 strain had the highest lovastatin yield, which was 23.93 μg/mL. The location of the tea forest strongly influenced the lovastatin yield of loose dark tea. The strain bred in this study improved the lovastatin yield of loose dark tea by more than three times when compared to wild Eurotium cristatum. These results are promising for the development of tea forest resources. [ABSTRACT FROM AUTHOR]

Dark tea (DT), a tea post-fermented by the Eurotium cristatum fungus, is known to attenuate oxidative damage and chronic inflammation. It also extends the lifespan of Caenorhabditis elegans fed a high-sugar diet. However, its impact on age-related muscle loss in mice is unclear. We divided 65-week-old male C57BL/6 mice into three groups: old control received the AIN-93 M diet, while the 0.15DT and 0.3DT groups had 0.15 % and 0.3 % DT for 9 weeks. As a result, DT improved grip strength, treadmill performance, muscle weight, and size in aged mice. It reduced cellular senescence, muscle atrophy markers, and angiopoietin-like 2 expression, while increasing sirtuin 1 protein levels in aged mice. Additionally, DT lowered malondialdehyde content and pro-inflammatory cytokine levels and enhanced total antioxidant capacity and enzymes, ultimately attenuating mitochondrial dysfunction in skeletal muscle. Therefore, our results suggest DT attenuates age-related muscle atrophy by alleviating oxidative stress, mitochondrial dysfunction, and inflammation.

Tea is one of the most popular drinks in the world. Dark tea is a kind of post-fermented tea with unique sensory characteristics that is produced by the special fermentation of microorganisms. It contains many bioactive substances, such as tea polyphenols, theabrownin, tea polysaccharides, etc., which have been reported to be beneficial to human health. This paper reviewed the latest research on dark tea's potential in preventing and managing cancer, and the mechanisms mainly involved anti-oxidation, anti-inflammation, inhibiting cancer cell proliferation, inducing cancer cell apoptosis, inhibiting tumor metastasis, and regulating intestinal flora. The purpose of this review is to accumulate evidence on the anti-cancer effects of dark tea, the corresponding mechanisms and limitations of dark tea for cancer prevention and management, the future prospects, and demanding questions about dark tea's possible contributions as an anti-cancer adjuvant. [ABSTRACT FROM AUTHOR]

Dark tea has great potential in regulating glycolipid metabolism, and theabrownin (TB) is considered to be the characteristic and bioactive constituent of dark tea. This study evaluated the ability of TB1 (fermented for 7 days) and TB2 (fermented for 14 days) isolated from dark tea to reverse insulin resistance (IR) in HepG2 cells. The results indicated that TB significantly ameliorated oxidative stress by improving mitochondrial function. In addition, TB improved glycogen synthesis and glucose consumption, and inhibited gluconeogenesis and fatty acid synthesis, by regulating GSK3β (Glycogen synthase kinase 3β), G6Pase (Glucose-6-phosphatase), GCK (Glucokinase), PEPCK1 (Phosphoenolpyruvate carboxy kinase 1), SREBP-1C (sterol regulatory element-binding protein 1C), FASN (fatty acid synthase), and ACC (Acetyl-CoA carboxylase). Additionally, the results of Western blot and real-time PCR experiments demonstrated that TB modulated glucolipid metabolism through the IRS-1 (Insulin receptor substrate 1)/PI3K (phosphatidylinositol-3 kinase)/Akt (protein kinase B) signaling pathway. Treatment with the PI3K inhibitor demonstrated a favorable correlation between PI3K activation and TB action on glycolipid metabolism. Notably, we observed that TB2 had a greater effect on improving insulin resistance compared with TB1, which, due to its prolonged fermentation time, increased the degree of oxidative polymerization of TB. [ABSTRACT FROM AUTHOR]

Ishizuchi dark tea, a traditional Japanese post-fermented tea, has attracted considerable attention for its bioactivity. In this study, the effect of water hardness on Ishizuchi dark tea infusions was analyzed. Sensory testing revealed that the teas brewed in hard water were less sour in taste than those brewed in soft water. The most abundant organic acid found in the tea infusions was lactic acid, with a constant concentration of 4 mmol L−1, regardless of water hardness. The pH of the tea infusions increased from 4 to 5 with increasing water hardness, and a similar increase in pH was observed with increasing concentrations of Ca2+ (or Mg2+) and HCO32− in an aqueous solution of 4 mmol L−1 lactic acid. The antioxidative activity of the tea infusion was clearly independent of water hardness. Therefore, hard water is suitable for brewing Ishizuchi dark tea because tea brewed in hard water has a less sour taste, which is more favored, while maintaining the same degree of antioxidative ability.

In this study, the solid-state fermentation (SSF) of dark tea was carried out using Bacillus subtilis LK-1, which was isolated from Fu brick tea (FBT). The effects of SSF with B. subtilis on volatile organic compounds (VOCs), non-volatile metabolites, and antioxidant activities of dark tea was investigated. A total of 45 VOCs were identified, primarily consisting of ketones (18), hydrocarbons (8), aldehydes (7), and alcohols (6). Following fermentation, the content of key odor active substances such as linalool, β-ionone, and 3,5-octadiene-2-one significantly increased, resulting in an enhanced floral and fruity aroma of dark tea. Furthermore, new flavor substances like geranyl isovalerate and decanal were produced during SSF, enriching the aroma profile of dark tea. Non-ester catechins demonstrated a drastic increase, while ester catechins remarkably decreased after SSF. Furthermore, SSF led to a slight decrease in the total polyphenols content and antioxidant activity of dark tea. There is a close relationship between VOCs and the main non-volatile metabolites during SSF. Overall, this study highlighted the great impact of SSF with B. subtilis on the metabolites of dark tea and provided valuable insights into the role of bacteria in shaping the metabolite profile of FBT.

Microorganisms in dark tea play an indispensable role in its ecosystem management. Despite the challenging conditions inherent to dark tea, a mere gram of its matrix can harbor a diverse array of microbial taxa, including bacteria and fungi. The interplay between dark tea and its microbiome is essential for ensuring quality and safety. Beneficial microbial communities significantly contribute to dark tea's rheological properties and organoleptic characteristics. This paper aims to elaborate on microbiome ecosystems in dark tea, microbial functions on dark tea quality, technology advances in microbiome research, and challenges of revealing microbiota-dark tea substrate interactions. The dark tea industry is now well-positioned to identify how to manipulate and manage the dark tea microbiome to enhance flavor, ensure safety, and improve our understanding of the mechanisms by which these ecosystems will adapt to environmental changes. • The structure, origin, and function of dark tea microbiome were introduced. • Integrated methods for zooming the diversity of dark tea microbiomes were introduced. • The new biotechnologies can be applied to tailor microbes for dark tea flavor. [ABSTRACT FROM AUTHOR]

The fermentation is the main process to form the unique flavor and health benefits of dark tea. Numerous studies have indicated that the microorganisms play a significant part in the fermentation process of dark tea. Dark tea has the quality of “The unique flavor grows over time,” but unscientific storage of dark tea might cause infestation of harmful microorganisms, thereby resulting in the remaining of fungi toxins. Mycotoxins are regarded as the main contributor to the quality of dark tea, and its potential mycotoxin risk has attracted people’s attention. This study reviews common and potential mycotoxins in dark tea and discusses the possible types of masked mycotoxins in dark tea. A summary of the potential risks of mycotoxins and masked mycotoxins in dark tea is presented, intending to provide a reference for the prevention and risk assessment of harmful fungi in dark tea. [ABSTRACT FROM AUTHOR]

Liupao tea is a postfermented dark tea with hypolipidemic activity. Research on the active substances in Liupao tea has primarily focused on those derived from the tea itself, overlooking the secondary metabolites produced by its predominant fungus, Euirotium cristatum . In this study, E. cristatum CPCC 401251, the predominant strain found in Liupao tea under investigation, was isolated and analyzed. A total of 19 representative metabolites, including prenylbenzaldehydes, diketopiperazines, and anthraquinones, were obtained from its culture. Subsequent analysis revealed the presence of multiple secondary metabolites of E. cristatum CPCC 401251 in Liupao tea. The 19 compounds significantly reduced the lipid content in free fatty acid (FFA)-stimulated hepatocyte AML-12 cells to varying degrees. Considering the content, chemical class, and biological activity of secondary metabolites from E. cristatum CPCC 401251, compounds 1 , 7 , and 13 were selected to detect their hypolipidemic activities and potential mechanisms in hyperlipidemia golden hamsters. Compound 1 exerted a hypolipidemic effect by activating the AMPK signaling pathway, decreasing Scd1, and improving intestinal flora. Compounds 7 and 13 played a role in the hypolipidemic activity by regulating the gene expression related to lipid synthesis and degradation, including upregulating the mRNA levels of Ppar α, Hsl , and Atgl , and decreasing the mRNA level of Scd1 . These findings help us understand the dominant fungus E. cristatum secondary metabolites presenting in Liupao tea and their potential hypolipidemic contributions. This work improves the understanding of the active substances in Liupao tea and highlights the health-promoting effects of microorganisms in the fermented tea.

Non-alcoholic fatty liver disease (NAFLD) has emerged as a leading public health challenge and is closely associated with metabolic syndromes, such as obesity. Intestinal microbiota dysbiosis could play a vital role in the pathogenesis and progression of NAFLD. Tea is the second most popular health drink in the world behind water, and exhibits many health-promoting effects. In this study, the protective effects of different black and dark teas on NAFLD induced by long-term high-fat diet (HFD) exposure and their regulation of gut microbiota were evaluated and explored. The results indicated that supplementation with different black and dark tea extracts could significantly suppress the energy intake, alleviate abnormal accumulation of visceral fat, and prevent obesity, hepatic abnormal lipid deposition and liver steatosis in HFD-fed mice at varying degrees. In addition, Dianhong tea and Liupao tea interventions could significantly decrease the ratio of Firmicutes to Bacteroidetes, and selenium-enriched black tea and selenium-enriched dark rea supplementation could remarkably reduce the relative abundance of Actinobacteria compared to the model group. Moreover, these teas could partly shift the relative abundances of Allobaculum, Roseburia and Dubosiella. Taken together, black teas and dark teas could prevent HFD-induced features of obesity and NAFLD, which might partly be due to the modulation of gut microbiota. [ABSTRACT FROM AUTHOR]

The epidemic of obesity is a serious public health problem. In this study, the effect of theabrownin from dark tea on obesity was evaluated by biochemical tests and nuclear magnetic resonance in C57BL/6J mice fed a high-fat diet. A mixture of antibiotics was used to deplete gut microbiota and then fecal microbiota transplant was used to restore gut microbiota. Untargeted metabolomics was used to reveal the effects of theabrownin on metabolic profiles through gut microbiota. The results showed that theabrownin significantly reduced body weight gain (83.0%) and body fat accumulation (30.29%) without affecting appetite. Also, theabrownin promoted lipid clearance with a hepatoprotective effect. The extra antibiotics disrupted the regulation of theabrownin on weight control while fecal microbiota transplant restored the beneficial regulation. That is, gut microbiota was important for theabrownin to reduce body weight gain. The untargeted metabolomics indicated that 18 metabolites were related to the anti-obesity effect of theabrownin mediated by gut microbiota. Furthermore, phenylalanine metabolism, histidine metabolism, as well as protein digestion and absorption pathway played a role in the anti-obesity of theabrownin. Our findings suggested that theabrownin significantly alleviated obesity via gut microbiota-related metabolic pathways, and theabrownin could be used for the prevention and treatment of obesity. [ABSTRACT FROM AUTHOR]

Background and aimsSichuan dark tea (ST), Zangcha, is a traditional fermented Chinese tea found in Sichuan and Tibet and claimed for beneficial effects against lifestyle-related metabolic disorders. We examined the effects of ST on lipid metabolism and atherosclerosis.Methods and resultsSichuan dark tea was given to fat-rich diet-induced atherosclerosis model rats in comparison with dark-fermented Chinese Pu-erh tea (PT) and Japanese green tea (GT). After 8 weeks of feeding, ST and PT induced an increase in high-density lipoprotein (HDL)-cholesterol and a decrease in glucose, and ST decreased triglyceride in plasma. ST also induced low pH in the cecum. There was no significant change in their body weight among the fat-feeding groups but a decrease was found in the visceral fat and liver weight in the ST group. Accordingly, ST reduced lipid deposition in the aorta in comparison with PT and GT. ST increased mRNA of LXRα, PPARα, PPARγ, and ABCA1 in the rat liver. The extract of ST stimulated the AMPK pathway to increase the expression of ABCA1 in J774 cells and increased expression of lipoprotein lipase and hormone-sensitive lipase in 3T3L1 cells, consistent with its anti-atherogenic effects in rats. High-performance liquid chromatography analysis showed unique spectra of original specific compounds of caffeine and catechins in each tea extract, but none of them was likely responsible for these effects.ConclusionSichuan dark tea increases plasma HDL and reduces plasma triglyceride to decrease atherosclerosis through AMPK activation. Further study is required to identify specific components for the effects of this tea preparation.

Glycolipid metabolic disorder is a serious threat to human health. Dark tea is a kind of traditional Chinese tea, which may regulate the glycolipid metabolic disorders. Dark tea extract (DTE) is the water extraction obtained from dark tea. Compared with traditional dark tea, DTE has the benefits of convenient consumption and greater potential for promoting health. However, the regulation of DTE on glycolipid metabolism and its molecular mechanism is rarely investigated. In our study, DTE was used as raw material to study the effect and molecular mechanism of its intervention on the glycolipid metabolic in db/db diabetic mice by using multiomics analysis and modern biological techniques. (1) DTE could significantly reduce fasting glucose in diabetic db/db mice, and the higher dose group has a better effect. Histopathological examination showed that DTE slightly improve the number of islets and decrease the number of islet β cells in the pancreatic tissue in db/db mice. (2) RNA-Seq was used to analyze the gene expression in liver tissue. In terms of biological processes, DTE mainly affected the inflammation and fatty acid metabolism. In terms of cell components, the lipoprotein and respiratory chain are mainly affected. In the aspect of molecular function, DTE mainly affected the redox related enzyme activity, iron ion binding and glutathione transferase. Arachidonic acid metabolism pathway, glutathione metabolism and PPAR signaling pathway were enriched by DTE with the results of KEGG pathway enrichment. In addition, real-time PCR results confirmed that DTE could significantly activate key genes of PPAR signaling pathway like Fabp1, Cyp4a1, Ehhadh, Cyp4a32, Aqp7 and Me1. (3) 16s rDNA showed that DTE could significantly decrease the ratio of Firmicutes/Bacteroidetes and the abundance of Proteobacteria, and increased the relative abundance of Verrucomicrobia at the phylum level. At the genus level, the relative abundance of Akkermansia, Prevotellaceae, Bacteroides and Alloprevotella was significantly increased after DTE treatment. This study provides multiomics molecular evidence for the intervention effect of DTE on abnormal glucose and lipid metabolism and the application of precise nutritional diet intervention of dark tea extract.

[Display omitted] • (1) The aroma compounds of dark teas from five different regions were thoroughly studied. • (2) A total of 1372 volatile compounds were detected in five regions dark teas. • (3) β-Ionone, dihydro-β-ionone, etc. compounds were identified as key aroma compounds. • (4) Eighteen aroma compounds can be used as factors to distinguish dark teas in PLS-DA. • (5) Five regions dark teas can be accurately categorized by Pearson correlation analysis. Dark tea (DT) holds a rich cultural history in China and has gained sizeable consumers due to its unique flavor and potential health benefits. In this study, headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC–MS), relative odor activity value (ROAV), and chemometrics approaches were used to detect and analyze aroma compounds differences among five dark teas from different geographical regions. The results revealed that the five DTs from different geographical regions differed in types, quantities, and relative concentrations of volatile compounds. A total of 1372 volatile compounds of were identified in the 56 DT samples by HS-SPME-GC–MS. Using ROAV and chemometrics approaches, based on ROAV>1 and VIP>1. Eighteen key aroma compounds can be used as potential indicators for DT classification, including dihydroactinidiolide, linalool, 1,2,3-trimethoxybenzene, geranyl acetone, 1,2,4-trimethoxybenzene, cedrol, 3,7-dimethyl-1,5,7-octatrien-3-ol, β -ionone, 4-ethyl-1,2-dimethoxybenzene, methyl salicylate, α -ionone, geraniol, linalool oxide I, linalool oxide II, 6-methyl-5-hepten-2-one, α -terpineol, 1,2,3-trimethoxy-5-methylbenzene, and 1,2-dimethoxybenzene. These compounds provide a certain theoretical basis for distinguishing the differences in five DTs from different geographical regions. This study provides a potential method for identifying the volatile substances in DTs and elucidating the differences in key aroma compounds. Abbreviations: DT, dark tea; FZT, Fuzhuan tea; LPT, Guangxi Liupao tea; QZT, Hubei Qingzhuan tea; TBT, Sichuan Tibetan tea; PET, Yunnan Pu-erh tea; ROAV, Relative odor activity value; OT, Odor threshold; HS-SPME, Headspace solid-phase microextraction; GC–MS, Gas chromatography-mass spectrometry; PCA, Principal components analysis; PLS-DA, Partial least squares-discriminant analysis; HCA, Hierarchical clustering analysis. [ABSTRACT FROM AUTHOR]

Aspergillus niger is one of the dominant microorganisms presented in dark tea fermentation. In this study, the biotransformation of steamed green tea leaves fermented by A. niger PW-2 was characterized using metabolomic and proteomic approaches. We observed that, after fermentation, the contents of volatile compounds contributing to the "green" aroma, including linalool, L-α-terpineol and geraniol, decreased significantly. Meanwhile, the astringency taste and contents of metabolites contributing to the taste (catechins) reduced significantly during fermentation. Additionally, the contents of theabrownins, which have health benefits, obviously increased. The bitter and umami tastes were also changed due to the variations in bitter-taste and umami-taste amino acids. We also found that glycoside hydrolases, tannases, catechol oxidases, peroxidases and laccases secreted by A. niger PW-2 were responsible for the metabolism of phenolic compounds and their derivatives (theaflavins, thearubingins and theabrownins). Finally, the metabolic pathways involved in the biosynthesis and degradation of characteristic metabolites were found to reveal the biotransformation characteristics of dark tea fermented with A. niger PW-2. [ABSTRACT FROM AUTHOR]

Instant dark tea (IDT) is a new product gaining increasing attention because it is convenient and can endow significant health benefit to consumers, which is partially attributed to its high concentration of functional ingredients. However, the molecular mechanism underlying its regulatory effect on hyperlipidaemia is rarely studied. In this study, we performed omics and molecular verification in high-fat diet (HFD)-fed rat, aiming to reveal the mechanism and provide molecular evidence. The results showed that the major bioactive components in IDT were 237.9 mg/g total polysaccharides, 336.6 mg/g total polyphenols, and 46.9 mg/g EGCG. Rats fed with IDT (0.27–0.54 g/kg for 12 weeks) significantly reduced the body weight and TC, TG, LDL-C, blood glucose, and MDA and induced the level of serum HDL-C and also the levels of liver SOD, CAT, GSH-Px, and Nrf2, compared to HFD group. For molecular mechanism study, HIDT feeding had significant impact on the gene expressions of biomarkers in lipogenesis (FABP, CD36, SCD1, Cyp4a1, and Kcnn2), lipid oxidation (PPARγ), and glucose glycolysis (Gck and ENO2) in liver tissue. Moreover, gut microbiome study found that rats fed with IDT dramatically modified the gut microbial species at the family level, such as suppressing the increase abundance of Proteobacteria and Firmicutes induced by HFD. HIDT significantly boosted the relative composition of beneficial bacterium Akkermansia and Rikenellaceae_RC9_gut_group and decreased the relative abundance of the harmful bacterium Ruminococcaceae_UCG-005 and Ruminiclostridium_9, compared to HFD (p < 0.01). Correlation analysis between microbiome and animal indicators found that seven genera including Akkermansia, Clostridiales, Lachnospiraceae, Lachnospiraceae_UCG-010, Ruminiclostridium_9, Ruminococaceae-UCG-005, and Ruminocuccus_1 were found as potential biomarkers that were strongly correlated with oxidative stress and metabolism genes. For instance, Ruminococcaceae_UCG-005 was significantly correlated with body weight, TG, HDL-C, Nfr2, FABP3, SCD1, Cyp4a1, and Kcnn2. Collectively, the above data obtained in this study had provided the primary molecular evidence for the molecular mechanism and brought in novel insights based on omics for the regulatory effect of IDT on hyperlipidaemia.

BACKGROUND: Dark tea, comprising one of the six major teas, has many biological activities, which originate from their active substrates, such as polyphenols, polysaccharides, and so on. The hypoglycemic effect is one of its most prominent activities, although less is known about their evaluation and potential role in the hypoglycemic mechanism. RESULTS: In the present study, we separately analyzed the phytochemical composition, glycosidase inhibition and free radical scavenging activities, and hypoglycemic activity in type 2 diabetes mellitus mice, as well as the alleviation of insulin resistance in HepG2 cells of four dark tea aqueous extracts. The results showed that the phytochemical composition of dark tea aqueous extracts was significantly different, and they all had good glycosidase inhibition and free radical scavenging activities, in vivo hypoglycemic activity and alleviation of insulin resistance, and could also activate the phosphatidylinositol 3‐kinase‐Akt‐perixisome proliferation‐activated receptor cascade signaling pathway to regulate glucose and lipid metabolism, change the key enzyme activities related to glucose metabolism and antioxidant activity, and reduce oxidative stress and inflammatory factor levels. Among them, Liubao brick tea (LBT) and Pu‐erh tea (PET) possessed better glycosidase inhibitory activity, in vivo hypoglycemic activity and improved insulin resistance activity, whereas Qingzhuan brick tea and Fuzhuan brick tea had better free radical scavenging activity, which may be explained by their distinct phytochemical compositions, such as tea proteins, polysaccharides, polyphenols, catechins, and tea pigments and some elements. CONCLUSION: Dark tea is a highly attractive candidate for developing antidiabetic food, LBT and PET may be good natural sources of agricultural products with anti‐diabetic effects. © 2021 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The present study aims to explore the glucose-lowering effects of the previously characterised dark tea (Camellia sinensis L.) protein extract (DTPE) from Heimaojian on the spleen–brain axis of diabetic mice. DTPE was orally administrated (50–100 mg/kg) to alloxan-induced mice for 21 d; a biochemical assay and transcriptome profiling (RNA sequencing (RNA-Seq)) were performed. The results showed that DTPE can improve glucose tolerance. Compared with the model group, at day 21, the fasting blood glucose values were significantly (P < 0·05) decreased by 44·9 % (13·8 v. 7·6 mmol/l) and 51·4 % (13·8 v. 6·7 mmol/l) for high dose of DTPE (100 mg/kg) and drug metformin (125 mg/kg) groups, respectively. Subsequently, transcriptome profiling (RNA-Seq) was performed on the spleen and brain of diabetic mice. Totally, fifty-two spleen-derived and forty-seven brain-derived differentially expressed genes related to the synthesis, transport and metabolism of glucose were identified. The regulatory network analysis indicated that DTPE may exert glucose-lowering effects through a thirty-seven-gene sub-network related to metabolism, Parkinson's disease, oxidative phosphorylation and immunity. In summary, for the first time, the present data revealed that dark tea-derived DTPE could exert a potential anti-hyperglycaemic effect by modulating the spleen–brain axis. [ABSTRACT FROM AUTHOR]

Theabrownins (TBs) are heterogeneous mixtures of water-soluble brown tea pigments, and important constituents to evaluate the quality of dark tea. TBs have numerous hydroxyl and carboxyl groups and are formed by the oxidative polymerization of tea polyphenols. Many biological activities attributed to TBs, including antioxidant, anti-obesity, and lipid-regulating, have been demonstrated. This review summarizes the research progress made on the formation mechanism and physicochemical properties of TBs. It also discusses their protective effects against various diseases and associated potential molecular mechanisms. Additionally, it examines the signaling pathways mediating the bioactivities of TBs and highlights the difficulties and challenges of TBs research as well as their research prospects and applications. [Display omitted] • Formation, preparation, and physicochemical characteristics of TBs were summarized. • The biological activity and mechanism of TBs were reviewed. • The challenges and strategies of current TB research are deeply analyzed. [ABSTRACT FROM AUTHOR]

The fungus Penicillium citrinum (P. citrinum), which is found in Chinese dark tea, is involved in the production of citrinin (CIT), a secondary metabolite with renal toxicity. In this study, a rapid method was used to detect CIT in Chinese dark tea samples from different regions and years. Of the 216 samples tested, 5 were detected to have CIT contamination, with a concentration ranging from 272.43 ± 16.57 to 649.68 ± 62.16 μg kg−1. Risk assessment for consuming dark tea showed that four heavy intake consumer groups in the deterministic risk assessment had hazard quotients (HQ) > 1, and the HQ for the probabilistic risk assessment was significantly below the acceptable level of 1.0. Furthermore, we found that tea polyphenols (TPs) inhibited the growth of P. citrinum. Transcriptome results showed that there were 483 downregulated and 783 upregulated genes between the TPs and control groups. Of which, orf1 and ctnD were significantly downregulated, demonstrating that TPs can affect CIT biosynthesis. Our research indicates that CIT in dark tea is within an acceptable risk level, and provides new insights into the inhibition of CIT. Furthermore, this study suggests that the suppression of CIT in dark tea by polyphenols make the consumption of dark tea safe. • Among all the samples tested, very few dark tea contained Citrinin (CIT). • The hazard quotients of consuming dark tea was below the acceptable level. • Tea polyphenols (TPs) inhibited the growth of Penicillium citrinum. • TP suppressed CIT biosynthesis by regulating the expression of related genes. [ABSTRACT FROM AUTHOR]

The latest insights into the health benefits of various fermented teas, including dark tea, oolong tea, and black tea, underscore the significant role of polyphenols in promoting wellness. The fermentation process in these teas not only enriches their overall polyphenol content but also enhances level of specific polyphenols, improving their health-promoting effects. Dark tea, such as Pu-erh, undergoes microbial fermentation, resulting in unique polyphenol profiles particularly theabrownins that offer potent antioxidant properties. Oolong tea, which is partially oxidized contains catechins and theasinensins, contributing to its potential to enhance weight management and correct metabolic disorders. Black tea, a fully oxidized tea, has a high levels of theaflavins and thearubigins, polyphenols linked to reduced inflammation, decreased incidences of cancer, and enhanced neuroprotection. However, despite these benefits, the emerging understanding of the health risks has associated fermented teas with the presence of potentially harmful compounds formed during fermentation or processing. In dark tea, prolonged microbial fermentation may lead to the production of mycotoxins, such as aflatoxins, which have been linked to carcinogenic effects. Black tea may accumulate heavy metals from soil and processing, posing risks to human health if consumed in excessive amounts. Similarly, oolong tea, while partially fermented, can still contain heavy metals contributing to impaired health. While these risks are generally low and depend on factors such as brewing conditions and consumption frequency, they highlight the importance of conducting future studies to devise preventive measures to mitigate these risks. [ABSTRACT FROM AUTHOR]

Fu Brick Tea (FBT), a distinguished dark tea, derives its unique sensory identity from a specialized fungal fermentation process. However, the effect of core functional fungi on sensory attributes during FBT fungal fermentation still needs to be systematically explored. Employing a synergistic blend of electronic sensory evaluation, microbiomics, and volatile metabolomics, this study delves into the dynamic evolution of fungi and volatile metabolites (VMs) during fungal fermentation. Utilizing fungal sequencing and Gas Chromatography-Mass Spectrometry (GC-MS) techniques, we unveiled 234 OTUs, with g_Eurotium emerging as a dominant fungus. Additionally, we identified 288 contributing VMs, 35 of which exhibited notable fluctuations during fermentation. These associated fungi influenced crucial metabolic pathways, enhancing the synthesis of sweet-scented components like acetophenone, while repressing woody and fecal aromas, exemplified by nerolidol, skatole, and indole. This study reveals the evolution of fungi and VMs during FBT fungal fermentation, and suggests 4 possible pathways of substance synthesis in which fungi may be involved, delivering valuable insights into the nuanced processes governing the quality formation of FBT. • g_Eurotium dominated the FBT fungal fermentation. • The fungal fermentation resulted in a distinctive aroma of green, floral, fresh, cucumber, and fruity. • Fungi modified synthetic pathways, enriching sweet aroma components while reducing woody, waxy, and fecal ones. [ABSTRACT FROM AUTHOR]

Non-alcoholic fatty liver disease (NAFLD) has emerged as a leading public health challenge and is closely associated with metabolic syndromes, such as obesity. Intestinal microbiota dysbiosis could play a vital role in the pathogenesis and progression of NAFLD. Tea is the second most popular health drink in the world behind water, and exhibits many health-promoting effects. In this study, the protective effects of different black and dark teas on NAFLD induced by long-term high-fat diet (HFD) exposure and their regulation of gut microbiota were evaluated and explored. The results indicated that supplementation with different black and dark tea extracts could significantly suppress the energy intake, alleviate abnormal accumulation of visceral fat, and prevent obesity, hepatic abnormal lipid deposition and liver steatosis in HFD-fed mice at varying degrees. In addition, Dianhong tea and Liupao tea interventions could significantly decrease the ratio of Firmicutes to Bacteroidetes, and selenium-enriched black tea and selenium-enriched dark rea supplementation could remarkably reduce the relative abundance of Actinobacteria compared to the model group. Moreover, these teas could partly shift the relative abundances of Allobaculum, Roseburia and Dubosiella. Taken together, black teas and dark teas could prevent HFD-induced features of obesity and NAFLD, which might partly be due to the modulation of gut microbiota.

The epidemic of obesity is a serious public health problem. In this study, the effect of theabrownin from dark tea on obesity was evaluated by biochemical tests and nuclear magnetic resonance in C57BL/6J mice fed a high-fat diet. A mixture of antibiotics was used to deplete gut microbiota and then fecal microbiota transplant was used to restore gut microbiota. Untargeted metabolomics was used to reveal the effects of theabrownin on metabolic profiles through gut microbiota. The results showed that theabrownin significantly reduced body weight gain (83.0%) and body fat accumulation (30.29%) without affecting appetite. Also, theabrownin promoted lipid clearance with a hepatoprotective effect. The extra antibiotics disrupted the regulation of theabrownin on weight control while fecal microbiota transplant restored the beneficial regulation. That is, gut microbiota was important for theabrownin to reduce body weight gain. The untargeted metabolomics indicated that 18 metabolites were related to the anti-obesity effect of theabrownin mediated by gut microbiota. Furthermore, phenylalanine metabolism, histidine metabolism, as well as protein digestion and absorption pathway played a role in the anti-obesity of theabrownin. Our findings suggested that theabrownin significantly alleviated obesity via gut microbiota-related metabolic pathways, and theabrownin could be used for the prevention and treatment of obesity.

Dark tea is a post-fermented tea with unique organoleptic characteristics. A unique microbial fermentation procedure is involved in dark tea processing that is considered a major factor to manage the quality of dark tea. Dark tea is gaining increasing attention recently because it contains various bioactive compounds with diverse biological functions. This review summarizes the microbial fermentation process, main bioactive compounds, and important biological functions or health benefits of dark tea, emphasizing its protective effects against different diseases and relevant molecular mechanisms, and proposes the perspectives in research and application of dark tea. Functional core microorganism-mediated bioconversion is crucial for the formation of dark tea's main quality components during the fermentation period. Dark tea contains a variety of bioactive compounds, including alkaloids, free amino acids, peptides, polyphenols, pigments, polysaccharides, and volatile compounds. Consumption of dark teas has been linked to multiple health benefits, such as antioxidant, anti-obesity, anti-diabetic, anti-cancer, cardiovascular protective, gastrointestinal protective, hepatoprotective, neuroprotective, and photoprotective activities. Future studies focusing on the health mechanisms of dark teas and the potential applications of dark tea in the food, pharmaceutical, and medical industries are guaranteed. • Dark teas are rich in diverse bioactive compounds. • Several core microorganisms are involved in dark tea fermentation. • Dark teas possess plenty of biological functions, especially anti-obesity and cardiovascular protective effects. • It is recommended for the public to consume dark teas to prevent and manage certain chronic diseases. [ABSTRACT FROM AUTHOR]

Aspergillus niger is one of the dominant microorganisms presented in dark tea fermentation. In this study, the biotransformation of steamed green tea leaves fermented by A. niger PW-2 was characterized using metabolomic and proteomic approaches. We observed that, after fermentation, the contents of volatile compounds contributing to the “green” aroma, including linalool, L-α-terpineol and geraniol, decreased significantly. Meanwhile, the astringency taste and contents of metabolites contributing to the taste (catechins) reduced significantly during fermentation. Additionally, the contents of theabrownins, which have health benefits, obviously increased. The bitter and umami tastes were also changed due to the variations in bitter-taste and umami-taste amino acids. We also found that glycoside hydrolases, tannases, catechol oxidases, peroxidases and laccases secreted by A. niger PW-2 were responsible for the metabolism of phenolic compounds and their derivatives (theaflavins, thearubingins and theabrownins). Finally, the metabolic pathways involved in the biosynthesis and degradation of characteristic metabolites were found to reveal the biotransformation characteristics of dark tea fermented with A. niger PW-2.

Objective: Eurotium sp. are the sexual states of the genus Aspergillus, and their ascospore is a spherical closed capsule with a golden colour. The growth of Eurotium sp. during tea production is a key step in achieving the unique quality of dark tea. This study aimed to evaluate the relationship between Eurotium sp. amount and Liupao tea quality. Methods and Results: The amounts of Eurotium sp. in 26 differently aged Liupao tea samples from several factories were studied. Indicators related to the quality of Liupao tea were investigated. The amounts of Eurotium sp. were divided into 0, 105 and 106 levels, and quantitative real‐time polymerase chain reaction (qPCR) was performed. Using ultrahigh‐performance liquid chromatography and high‐performance liquid chromatography, the amounts of emodin and physcion were determined to be closely related to the amount of Eurotium sp. Emodin was not found or occurred in minimal amounts in all raw Liupao tea samples. By contrast, physcion was found in Liupao tea at the 106 level of Eurotium sp. Liupao tea samples with varying levels of Eurotium sp. also exhibited evident differences in aroma and chromaticity. Result of the Pearson correlation test showed that the amount of Eurotium sp. plays a key role in creating the unique quality of Liupao tea. Conclusion: The amount of Eurotium sp. in dark tea detected via qPCR can be used as a quantitative quality indicator for evaluating dark tea. Significance and Impact of the Study: The present study provides an efficient method for identifying the different qualities of dark tea and addressing quality control issues in fermenting dark tea. [ABSTRACT FROM AUTHOR]

As the critical process for quality formation of ripened Pu-erh tea (RpPT), pile-fermentation mechanism has been widely studied by using omics approach, chemical analysis, microbial distribution and their inoculated fermentation tests. Despite several reviews about chemical compositions and health benefits in RpPT and other dark teas, more comprehensive study is requisite to summarize the pile-fermentation mechanism of RpPT and application prospects of relevant microbes. This paper aims to elaborate dominant microbes, recent advances in single- and multi-omics approaches, and pathways of characteristic components during the pile-fermentation. Microbial effects are summarized and compared, which would enhance their application potential in RpPT production. At least 93 fungi and 53 bacteria have been isolated from RpPT related to the pile-fermentation. Aspergillus niger is regarded as dominant microorganism in pile-fermentation for its higher detection frequency and relative abundance. Aspergillus , Penicillium, Rasamsoni, Rhizomucor and Bacillus genera promote RpPT quality formation. The low mycotoxins contamination risk in pile-fermentation and inoculated fermentation indicates microbial application potential in RpPT production. Several microbes are confirmed to promote formation of characteristic components including theabrownins, puerins, teadenol A, gallic acid, lovastatin, gamma-aminobutyric acid and several flavonols such as quercetin. Future studies focusing on superiority and application potential of relevant microbes in RpPT production through exogenous inoculation in the pile-fermentation are guaranteed, which could improve sensory quality and health benefits of RpPT, and shorten fermentation cycle. • A. niger dominates in pile-fermentation for quality formation of ripened Pu-erh tea (RpPT). • Low mycotoxins risk is confirmed both in pile-fermentation and inoculated fermentation. • Formation mechanisms of RpPT characteristics in pile-fermentation are clarified. • Several microbes promote formation of characteristic components such as theabrownins. • Microbial exogenous inoculation show application potential in RpPT production. [ABSTRACT FROM AUTHOR]

Sichuan dark brick tea (Camellia sinensis) and Sichuan Fuzhuan brick tea have significantly different aroma characteristics although both of them have almost the same processing methods. Thus, these two types of tea were used as the research materials to determine the differences in their aroma compounds. The volatile compounds in the two types of tea were identified and quantified by headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME–GC–MS), results showed that they both had 37 common volatile compounds. Then the aroma-active components were identified by odour activity value (OAV). It was found that SFBT had 20 aroma-active components, of which β-ionone had the largest OAV (199547.72). SDBT has 21 aroma-active ingredients (including all 20 aroma-active components of SFBT), of which β-ionone again has the largest OAV (114800.66). Finally, the aroma profile differences between the two tea samples were studied by aroma profile tests, and the results showed that the main aroma differences of SDBT and SFBT were caused by β-ionone, epoxydihydrolinalool II, methyl salicylate, geranylacetone, nerolidol, benzaldehyde, benzyl acetate, nonanal, trans,trans-2,4-heptadienal and 1-octen-3-ol, in addition, defined SFBT's 'fungi flower aroma' and SDBT's 'aged fragrance' from the level of aroma monomer. [ABSTRACT FROM AUTHOR]

Fluorosis caused by consumption of dark tea is a major public health problem in the western part of China. In this study, the effects of milk powder, butter, and table salt on the bioavailability of the fluoride in an infusion of dark tea were investigated in a rat model. These substances were examined for their potential as dietary interventions to reduce the risk of tea-induced fluorosis. Fluoride was less bioavailable from the dark tea infusion than from a NaF solution. The addition of milk powder significantly decreased the amount of fluoride absorbed from the dark tea infusion and increased the amount of fluoride in fecal excretion. While butter had a limited effect on fluoride bioavailability, it prolonged the fluoride absorption period. The addition of 4 mg/mL table salt significantly increased the bioavailability of fluoride in the dark tea infusion. The addition of different flavor augmentations to a dark tea infusion had different effects on fluoride bioavailability. Therefore, dietary intervention may be a novel strategy for reducing fluorosis risk. • The bioavailability of fluoride in dark tea and NaF aqueous solution was evaluated. • Milk significantly reduced the bioavailability of fluoride in dark tea. • In rats, butter prolonged the absorption period of fluoride from dark tea. [ABSTRACT FROM AUTHOR]

Abstract Dark tea is a special type of tea that requires microbial fermentation during the post-fermentation process, and its volatile composition is quite different from other tea kinds. The present study aimed at charactering the volatile composition of two typical Chinese dark teas (Fu-brick tea and Pu-erh tea). Volatile compounds from both teas were extracted and analyzed using simultaneous distillation extraction (SDE) combined with two-dimensional gas chromatography-time-of-flight-mass spectrometry (GC × GC-TOFMS). A total of 373 and 408 major aromatic components were tentatively identified in Fu-brick tea and Pu-erh tea, respectively. The relative quantification results showed that ketenes (24.87%), ketones (16.86%), aldehydes (14.36%) and olefine aldehydes (9.11%) were more abundant in Fu-brick teas, while in Pu-erh teas, ketenes accounted for 17.95%, followed by aldehydes (14.51%), ally esters and ketones (11.94% and 10.78%). Multivariate analysis indicated that obvious differences existed in the levels of some aromatic compounds between the two dark teas. Benzaldehyde and benzeneacetaldehyde in Fu-brick teas were considerably lower and accounted for approximately 55% of those in Pu-erh teas; whilst, nonanal and 2-hexenal in Fu-brick tea were twice as high as in Pu-erh tea. The application of pre-enrichment extraction with GC × GC-TOFMS analysis gives a thorough view of the aromatic constituents of dark teas. Graphical abstract Image 1 Highlights • Fu-brick and Pu-erh teas were compared using SDE-GC × GC-TOF/MS. • Ketenes were the most abundant aroma components in both dark teas. • Concentrations of nonanal and 2-hexenal in Fu-brick were twice those in Pu-erh tea. • This is the first comprehensive report of volatile compounds in two dark teas. [ABSTRACT FROM AUTHOR]

[Display omitted] • Dark tea (DT) improved the progressive loss of motor control in aged mice. • DT ameliorated age-related skeletal muscle atrophy in skeletal muscle of mice. • DT suppressed oxidative stress and enhanced antioxidant capacity and mitochondrial function in mice. • DT alleviated inflammation in skeletal muscle of aged mice. Dark tea (DT), a tea post-fermented by the Eurotium cristatum fungus, is known to attenuate oxidative damage and chronic inflammation. It also extends the lifespan of Caenorhabditis elegans fed a high-sugar diet. However, its impact on age-related muscle loss in mice is unclear. We divided 65-week-old male C57BL/6 mice into three groups: old control received the AIN-93 M diet, while the 0.15DT and 0.3DT groups had 0.15 % and 0.3 % DT for 9 weeks. As a result, DT improved grip strength, treadmill performance, muscle weight, and size in aged mice. It reduced cellular senescence, muscle atrophy markers, and angiopoietin-like 2 expression, while increasing sirtuin 1 protein levels in aged mice. Additionally, DT lowered malondialdehyde content and pro-inflammatory cytokine levels and enhanced total antioxidant capacity and enzymes, ultimately attenuating mitochondrial dysfunction in skeletal muscle. Therefore, our results suggest DT attenuates age-related muscle atrophy by alleviating oxidative stress, mitochondrial dysfunction, and inflammation. [ABSTRACT FROM AUTHOR]

The pile fermentation process of Fuzhuan brick tea is unique in that it involves preheating without the use of starter cultures. The detailed metabolite changes and their drivers during this procedure are not known. Characterizing these unknown changes that occur in the metabolites and microbes during pile fermentation of Fuzhuan brick tea is important for industrial modernization of this traditional fermented food. Using microbial DNA amplicon sequencing, mass spectrometry-based untargeted metabolomics, and feature-based molecular networking, we herein reveal that significant changes in the microbial community occur before changes in the metabolite profile. These changes were characterized by a decrease in Klebsiella and Aspergillus , alongside an increase in Bacillus and Eurotium . The decrease in lysophosphatidylcholines, unsaturated fatty acids, and some astringent flavan-3-ols and bitter amino acids, as well as the increase in some less astringent flavan-3-ols and sweet or umami amino acids, contributed importantly to the overall changes observed in the metabolite profile. The majority of these changes was caused by bacterial metabolism and the corresponding heat generated by it.

The contributions of four kinds of dark tea (Camellia sinensis L.) proteins and their hydrolysates to hypoglycemic activity were investigated in vitro. Four kinds of water-extracted dark tea proteins were hydrolyzed with trypsin and Alcalase, respectively. The complete proteins had α-amylase inhibitory activity with half maximal inhibitory concentration (IC50) values ranging from 1.27 to 2.78 mg/mL. Most of the dark tea proteins and hydrolysates significantly inhibited α-glucosidase and dipeptidyl peptidase (DPP-IV), with IC50 values in the range of 0.0103-1.3114 mg/mL and 0.1000-1.3364 mg/mL, respectively. In general, Heimaojian (HMJ) and Qianliang (QL) hydrolysates displayed high α-glucosidase inhibitory activity, while HMJ, Fuzhuan (FZ), and Heizhuan (HZ) hydrolysates exhibited a strong ability to inhibit DPP-IV. This study demonstrates the potential of dark tea proteins and their hydrolysates as a source of functional food and medicine for the control of type 2 diabetes. [ABSTRACT FROM AUTHOR]

Summary: Polysaccharides‐rich dark tea is believed to be beneficial to health in Southeast Asian. However, the hypolipidemic effect of Chinese Liupao tea polysaccharides (CLTPS) has not been investigated. Here, the potential mechanism by which the oral administration of CLTPS affects lipid metabolism was evaluated using high‐fat diet‐induced hyperlipidemia rats. In rats supplemented with CLTPS, a suppressive effect on body weight was observed after 4 weeks of intervention. CLTPS significantly improved the levels of lipid profiles, the oxidation of lipids and antioxidant enzyme activity in a dose‐dependent manner. The cholesterol‐lowering action was paralleled by the stimulation of cholesterol converted to bile acids along with a 1.5‐fold increase in faecal excretion. Besides, the prolongation of coagulation factors and the reduction in fibrinogen content showed that CLTPS regulated the coagulation initiated through the intrinsic pathway. With current findings, CLTPS may be valuable as a natural hypolipidemic nutraceutical for human consumption. [ABSTRACT FROM AUTHOR]

• Widely targeted metabolomics reveals chemical periodic change in whole process. • 568 Differential metabolites (VIP > 1.0, P < 0.05, FC > 1.50 or < 0.67) selected. • Anaerobic fermentation is regarded as critical process for the quality formation. • 7 Amino acids, 4 phenolic acids, 6 flavonoids and their glycosides accumulated. • N -, O -methylation, hydrolyzation, glycosylation, oxidation involved in fermentation. Yunnan pickled tea is produced from fresh tea-leaves through fixation, rolling, anaerobic fermentation and sun-drying. In this study, widely targeted metabolomics using UHPLC-QQQ-MS/MS and HPLC analysis were carried out to elaborate its quality formation during the whole process. Results confirmed the contribution of preliminary treatments and anaerobic fermentation to the quality formation. A total of 568 differential metabolites (VIP > 1.0, P < 0.05, FC > 1.50 or < 0.67) were screened through OPLS-DA. (-)-Epigallocatechin and (-)-epicatechin significantly (P < 0.05) increased from the hydrolyzation of ester catechins, such as (-)-epigallocatechin gallate and (-)-epicatechin gallate in anaerobic fermentation. Additionally, the anaerobic fermentation promoted vast accumulations of seven essential amino acids, four phenolic acids, three flavones and flavone glycosides, pelargonidin and pelargonidin glycosides, flavonoids and flavonoid glycosides (i.e. kaempferol, quercetin, taxifolin, apigenin, myricetin, luteolin and their glycosides) through relevant N -methylation, O -methylation, hydrolyzation, glycosylation and oxidation. [ABSTRACT FROM AUTHOR]