Aspergillus Cristatus Research Articles

Screening Proteins That Interact With AcHog1 and the Functional Analysis of AcSko1 in Aspergillus cristatus.

Aspergillus cristatus is a dominant fungus formed during the "flowering" process of Fuzhuan brick tea. Previous research has established that the sporulation of Aspergillus nidulans, a model organism of filamentous fungi, is regulated by light. However, the sporulation of A. cristatus is dependent on osmotic stress. In a previous study, we used pull-down and mass spectrometry to identify proteins that interacted with AcHog1 in A. cristatus when cultured under different conditions of osmotic stress. In the present study, we analyzed the proteins we identified previously to investigate their functional role. The AA1E3BER4 protein was located downstream of Hog1 in the HOG branch pathway and was identified that was regulated by AcHog1. Furthermore, yeast two-hybrid analysis showed that AA1E3BER4 interacted with AcHog1. In addition, we knocked out and complemented the Acsko1 gene encoding the AA1E3BER4 protein. We found that the number of sexual and asexual spores were downregulated by 3.81- and 4.57-fold, respectively, in the ΔAcsko1 strain. The sensitivity of the ΔAcsko1 strain to sorbitol and sucrose, as regulators of osmotic stress, increased, and the sensitivity to high sucrose was higher than that of sorbitol. Acsko1 also regulated the response of A. cristatus to oxidative stress, Congo red, and SDS (sodium dodecyl sulfate). In addition, the deletion of Acsko1 significantly increased the pigment of the ΔAcsko1 strain. This is the first study to report the role of the sko1 gene in oxidative stress, stress-induced damage to the cell wall, and pigment in Aspergillus cristatus.

A special “Golden Flower” fungus with yellow cleistothecium: Its impact on the flavor of golden flower loose tea produced from summer-autumn tea

Summer-autumn tea leaves are abundant, yet their utilization remains low. Our study enhances the quality and flavor of Golden Flower loose tea (GFLT) from fresh summer-autumn tea leaves by using a special “Golden Flower” fungus (strain ACF-2). The strain ACF-2, identified as Aspergillus cristatus with yellow cleistothecium and isolated from Fu Brick Tea, was characterized by colony morphology, microstructural analyses, and phylogenetic examination using a 3-gene dataset (BenA, CaM, RPB2). Inoculation of raw dark tea products with A. cristatus led to a significant increase in tea water extract and a reduction in tea polyphenols, soluble sugars, flavonoids, and total amino acids, thereby enhancing the quality and flavor of GFLT. HS-SPME-GC-MS analysis of GFLT aroma revealed that A. cristatus substantially improved the tea’s aroma profile. 9 volatile compounds—(Z)-jasmone I, β-cyclocitral, linalool oxide I, linalool, hexanal, 1-octen-3-ol, (Z)-citral, citral, and methyl salicylate, were found to be significantly elevated in GFLT compared to the controls. Our findings provide both theoretical and practical insights into optimizing the utilization of summer-autumn tea leaves, identifying the “Golden Flower” fungus, and understanding its impact on GFLT quality. In summary, fermenting A. cristatus to produce GFLT may be a new channel to utilize summer-autumn tea.

Acptp2,3 participates in the regulation of spore production, stress response, and pigments synthesis in Aspergillus cirstatus.

Aspergillus cristatus was a filamentous fungus that produced sexual spores under hypotonic stress and asexual spores under hypertonic stress. It could be useful for understanding filamentous fungi's sporulation mechanism. Previously, we conducted functional studies on Achog1, which regulated the hyperosmotic glycerol signaling (HOG) pathway and found that SI65_02513 was significantly downregulated in the transcriptomics data of ΔAchog1 knockout strain. This gene was located at multiple locations in the HOG pathway, indicating that it might play an important role in the HOG pathway of A. cristatus. Furthermore, the function of this gene had not been identified in Aspergillus fungi, necessitating further investigation. This gene's conserved domain study revealed that it has the same protein tyrosine phosphatases (PTPs) functional domain as Saccharomyces cerevisiae, hence SI65_02513 was named Acptp2,3. The function of this gene was mostly validated using gene knockout and gene complementation approaches. Knockout strains exhibited sexual and asexual development, as well as pigments synthesis. Morphological observations of the knockout strain were carried out under several stress conditions (osmotic stress, oxidative stress, Congo Red, and sodium dodecyl sulfate (SDS). Real-time fluorescence polymerase chain reaction (PCR) identified the expression of genes involved in sporulation, stress response, and pigments synthesis. The deletion of Acptp2,3 reduced sexual and asexual spore production by 4.4 and 4.6 times, demonstrating that Acptp2,3 positively regulated the sporulation of A. cristatus. The sensitivity tests to osmotic stress revealed that ΔAcptp2,3 strains did not respond to sorbitol-induced osmotic stress. However, ΔAcptp2.3 strains grew considerably slower than the wild type in high concentration sucrose medium. The ΔAcptp2,3 strains grew slower than the wild type on media containing hydrogen peroxide, Congo red, and SDS. These findings showed that Acptp2,3 favorably controlled osmotic stress, oxidative stress, and cell wall-damaging chemical stress in A. cristatus. Deleting Acptp2,3 resulted in a deeper colony color, demonstrating that Apctp2,3 regulated pigment synthesis in A. cistatus. The expression levels of numerous stress-and pigments-related genes matched the phenotypic data. According to our findings, Acptp2,3 played an important role in the regulation of sporulation, stress response, and pigments synthesis in A. cristatus. This was the first study on the function of PTPs in Aspergillus fungi.

Constituent-taste relationship of Kuding tea fermented by Aspergillus neoniger and Aspergillus cristatus: Unveiling taste characteristics through untargeted metabolomics

Kuding tea (KDT), known for its medicinal and edible attributes, currently faces limited application due to its unfavorable taste. Thus, we aimed to improve KDT flavor using fungal fermentation and elucidate the mechanism of flavor change through the constituent-taste relationship. In this study, an electronic tongue was utilized to assess the flavor profiles of unfermented and fermented teas by Aspergillus neoniger (AN) and Aspergillus cristatus (AC). A constituent-taste relationship method combining untargeted metabolomics and taste sensors was employed to analyze the taste-related differential metabolites. The analysis of metabolic pathways further elucidated the potential taste-altering mechanisms. Fermentation of both AN and AC led to changes in the umami and sweetness of KDT. Additionally, AN fermentation impacted bitterness and sourness, while AC fermentation influenced saltiness. Based on the strategy of constituent-taste relationship, 31 and 30 metabolites predominantly including lipids, phenolic acids, flavonoids, amino acids and phenolic glycosides were discovered to be significantly correlated with the flavor characteristics of KDT. During fermentation, AN and AC participated in 5 and 4 metabolic pathways, respectively, showing distinct regulatory mechanisms in altering KDT flavor. AN led to pronounced deglycosylation of glycosides, while AC impacted the degradation of phenolic acids and lipids. Together, this study advances the understanding of metabolic changes in fermented KDT and provides valuable insights into the enhancement of flavor characteristics.

Bioturbation effect of high - Yield pyrazine strain on the microbial community and flavour metabolites of fortified Daqu

In order to comprehensively explore the regulatory effect of bioturbation on the assembly and overall metabolic characteristics of Daqu microbial community. This study used metatranscriptomics and metabolomics to investigate the composition of the microbial community, physicochemical properties, and overall metabolic profile of Daqu before and after bioturbation by Bacillus velezensis. The results demonstrated that the acidity, moisture, amino acid nitrogen, and starch content of Daqu exhibited a significant increase following bioturbation. The symbol microorganisms of Daqu have also changed significantly, from the original Aspergillus chevalieri, Rasamsonia emersonii, Aspergillus ruber, Aspergillus glaucus, Aspergillus wentii and Aspergillus cristatus to Paecilomyces varioti, Trichomonascus ciferrii and Staphylococcus gallinarum. Correlation analyses and integration pathways indicated that altered microbial community abundance also promoted the metabolism of starch, ethanol, fatty acids, and pyrazine, resulting in a significant enhancement of Daqu's saccharification and liquefaction power, as well as the content of some volatile and non-volatile metabolites, with pyrazines increased by 24.50%. The application of fortified Daqu to Baijiu fermentation can significantly enhance the content of various pyrazines in Baijiu. These results are helpful to elucidate the mechanism of biochemical changes, microbial assembly and flavor production of Daqu after biodisturbance.

Mechanisms of single and mixed microbial fermentation to improve summer-autumn green tea

To investigate the effect of individual and collaborative fermentation on summer-autumn green tea (SAGT) quality. Liquid fermentation with individual inoculation of Aspergillus cristatus (ACFT), Aspergillus niger (ANFT) and mixed inoculation of A. cristatus/A. niger (MFT), respectively, with SAGT extracts (CT) as raw materials. The volatile organic compounds (VOCs), characteristic non-volatile compounds and sensory evaluation of tea samples were determined. The results showed a significant decrease in the content of tea polyphenols, carbohydrates and nongalloylated catechins (C, EC) and a significant increase in the amount of VOCs in MFT compared to ANFT and ACFT. The results of metabolic pathway analysis and sensory evaluation showed that ACFT went through the terpene and phenylpropanoic acid metabolic pathway, which produced large amounts of α-terpineol, phenethyl alcohol, and methyl salicylate, which in turn conferred ACFT with distinctly sweet, floral, and herbal aroma. ANFT produces large amounts of linalool oxides I and II, mainly through the linalool oxide metabolic pathway, making ANFT a distinctive aged and woody aroma. MFT may be due to the strong woody aroma of linalool oxides masking other weaker and better aromas, thus giving MFT a significant woody, herbal and stale aroma. This study provides new insights and a theoretical basis for mixed fungal fermentation to improve the quality of SAGT.

Morphological variation and expressed sequence tags-simple sequence repeats-based genetic diversity of Aspergillus cristatus in Chinese dark tea.

Aspergillus cristatus is a homothallic fungus that is used in the natural fermentation process of Chinese Fuzhuan tea and has been linked to the production of bioactive components. However, not much is known about the variations present in the fungus. To understand the variation of the dominant microorganism, A. cristatus, within dark tea, the present study investigated the genetic and morphological diversity of 70 A. cristatus collected across six provinces of China. Expressed sequence tags-simple sequence repeats (EST-SSR) loci for A. cristatus were identified and corresponding primers were developed. Subsequently, 15 specimens were selected for PCR amplification. The phylogenetic tree obtained revealed four distinct clusters with a genetic similarity coefficient of 0.983, corresponding to previously identified morphological groups. Five strains (A1, A11, B1, D1, and JH1805) with considerable differences in EST-SSR results were selected for further physiological variation investigation. Microstructural examinations revealed no apparent differentiation among the representative strains. However, colony morphology under a range of culture media varied substantially between strains, as did the extracellular enzymatic activity (cellulase, pectinase, protease, and polyphenol oxidase); the data indicate that there are differences in physiological metabolic capacity among A. cristatus strains. Notably, JH1805, B1, and A11 exhibited higher enzymatic activity, indicating their potential application in the production of genetically improved strains. The findings provide valuable insights into species identification, genetic diversity determination, and marker-assisted breeding strategies for A. cristatus.

Research on the mechanism of sea buckthorn leaf Fu tea in the treatment of hyperlipidemia

BackgroundHyperlipidemia (HLP) presents a significant challenge to global public health. Mounting evidence suggests that statins, the recommended first-line lipid-lowering agents, have significant adverse effects. Consequently, the quest for natural and efficacious alternative therapies is steadily emerging as a research priority for HLP prevention and treatment. Consumption of tea, which is rich in diverse biologically active compounds with the capacity to regulate lipid metabolism and combat obesity, has emerged as a promising alternative therapy. Sea buckthorn leaves are rich in a multitude of biologically active substances, have a hypolipidemic effect, and can be used as a raw material for tea because of their unique flavor. There is a suggestion that combining Aspergillus cristatus with tea could modify or boost the lipid-lowering active compounds present in tea, thereby increasing its efficacy in regulating lipid metabolism. ResultsSea Buckthorn Leaf Fu Tea (SBLFT) was obtained by fermentation when sea buckthorn leaves contained 42 % moisture, inoculated with Aspergillus cristatus 0.2 mL/g, and incubated for 8 d at constant temperature. Animal experiments demonstrated that SBLFT significantly inhibited body weight gain in HLP rats and reduced lipid content and serum oxidative stress. In addition, liver tissue sections and functional indices showed that SBLFT can improve liver morphology and function abnormalities. Reverse transcription-polymerase chain reaction results indicated that the expression of Liver kinase B1 (LKB1), adenosine 5‘-monophosphate (AMP)-activated protein kinase (AMPK), acetyl CoA carboxylase 1 (ACC1), and sterol-regulatory element binding protein-1 (SREBP1c) gene related to lipid metabolism was altered. ConclusionSBLFT improved HLP, specifically via promoting the expression of LKB1 in the liver of HLP rats, activating AMPK, and inhibiting ACC1 and SREBP1c expression, resulting in the inhibition of fatty acid and triglyceride synthesis-related enzymes at the transcriptional level.

Effects of solid-state fermentation with Aspergillus cristatus (MK346334) on the dynamics changes in the chemical and flavor profile of dark tea by HS-SPME-GC–MS, HS-GC-IMS and electronic nose

Aspergillus cristatus, the predominant microbe of Fuzhuan brick tea (FBT), is responsible for the creation of distinctive golden flower and unique floral aroma of FBT. The present study examined the alterations in chemical and aromatic components of raw dark tea by solid-state fermentation using A. cristatus (MK346334), the strain isolated from FBT. As results, catechins, total ployphenols, total flavonoids, theaflavins, thearubigins and antioxidant activity were significantly reduced after fermentation. Moreover, 112 and 76 volatile substances were identified by HS-SPME-GC–MS and HS-GC-IMS, respectively, primarily composed of alcohols, ketones, esters and aldehydes. Furthermore, the calculation of odor activity values revealed that 19 volatile chemicals, including hexanal, heptanal, linalool and methyl salicylate, were the main contributors to the floral, fungal, woody and minty aroma of dark tea. The present research highlights the pivotal role played by the fermentation with A. cristatus in the chemical composition, antioxidant property and distinctive flavor of dark tea.

Material Composition Characteristics of Aspergillus cristatus under High Salt Stress through LC-MS Metabolomics.

Aspergillus cristatus is a crucial edible fungus used in tea fermentation. In the industrial fermentation process, the fungus experiences a low to high osmotic pressure environment. To explore the law of material metabolism changes during osmotic pressure changes, NaCl was used here to construct different osmotic pressure environments. Liquid chromatography-mass spectrometry (LC-MS) combined with multivariate analysis was performed to analyze the distribution and composition of A. cristatus under different salt concentrations. At the same time, the in vitro antioxidant activity was evaluated. The LC-MS metabolomics analysis revealed significant differences between three A. cristatus mycelium samples grown on media with and without NaCl concentrations of 8% and 18%. The contents of gibberellin A3, A124, and prostaglandin A2 related to mycelial growth and those of arabitol and fructose-1,6-diphosphate related to osmotic pressure regulation were significantly reduced at high NaCl concentrations. The biosynthesis of energy-related pantothenol and pantothenic acid and antagonism-related fluvastatin, aflatoxin, and alternariol significantly increased at high NaCl concentrations. Several antioxidant capacities of A. cristatus mycelia were directly related to osmotic pressure and exhibited a significant downward trend with an increase in environmental osmotic pressure. The aforementioned results indicate that A. cristatus adapts to changes in salt concentration by adjusting their metabolite synthesis. At the same time, a unique set of strategies was developed to cope with high salt stress, including growth restriction, osmotic pressure balance, oxidative stress response, antioxidant defense, and survival competition.

Polysaccharides of Aspergillus cristatus attenuate obesity by regulating gut microbiota and gut microbiota-related metabolites

Golden-flower fungus, the only dominant microorganism determining the Fu-brick tea quality through fermentation and the important microbe in Liupao tea, is considered a potential probiotic fungus based on its anti-obesity effect. However, the classification of golden-flower fungi is still controversial; the anti-obesity effect of golden-flower fungus polysaccharides remains unknown. In this study, we identify a golden-flower strain as Aspergillus cristatus based on morphological characteristics and multigene phylogeny analysis, which resolves the controversy of classification. Moreover, we find A. cristatus polysaccharides (ACPS) attenuate obesity in rats. ACPS modulate gut bacterial composition, in which Akkermansia, Akkermansia muciniphila, Bacteroides, Romboutsia, Blautia, and Desulfovibrio are considered the core microbes regulated by ACPS. ACPS increase fecal total short-chain fatty acid content and serum, hepatic, and fecal total bile acid content. Furthermore, ACPS-induced gut microbiota alteration plays a causal role in the protection from obesity, according to a fecal transplantation experiment. Thus, ACPS ameliorate obesity by regulating gut microbiota and gut microbiota-related metabolites.

Reexamination of Aspergillus cristatus phylogeny in dark tea: Characteristics of the mitochondrial genome.

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.

Aspercitrininone A, novel antibacterial polyketide featuring unusual spiral skeleton from Aspergillus cristatus

Aspercitrininone A (1), a novel polyketide featuring an unprecedented tetracyclic 6/6/6/5 spiral skeleton, was obtained from the rice fermentation cultures of the fungus Aspergillus cristatus together with five known compounds (2–6). Their structures were determined by HRESIMS data, 1D and 2D NMR spectroscopic analysis, and electronic circular dichroism (ECD) calculations. Aspercitrininone A was revealed as a new type of C/D cycle spiral structure and an unusual addition product of o-quinoid form citrinin with 2-methylterrefuranone. Compounds 1, 4, and 5 exhibited potent antibacterial activities with minimal inhibitory concentration (MIC) values from 13.2 to 67.3 μg/mL against four strains of human pathogenic bacteria in vitro.

Characteristic volatiles of Fu brick tea formed primarily by extracellular enzymes during Aspergillus cristatus fermentation

Fu brick tea (FBT) has unique “fungal flower” aroma traits, but its source of crucial aroma compounds is still controversial. Aspergillus cristatus is the dominant fungus that participated in the fermentation of FBT. In this study, volatiles of Aspergillus cristatus and corresponding fermented FBT were examined using GC × GC-Q-TOFMS. A total of 59 volatiles were shared by three strains of Aspergillus cristatus isolated from representative FBT. Among them, 1-octen-3-ol and 3-octanone were the most abundant. A total of 133 volatiles were screened as typical FBT volatiles from three FBTs fermented by the corresponding fungi. Aspergillus cristatus and FBT had only 29 coexisting volatiles, indicating that the volatiles of Aspergillus cristatus could not directly contribute to the aroma of FBT. The results of no significant correlation between volatile content in FBT and volatile content in Aspergillus cristatus suggested that intracellular metabolism of Aspergillus cristatus was not a direct driver of FBT aroma formation. Metabolic pathway analysis and proteomic analysis showed that the aroma in FBT was mainly formed by the enzymatic reaction of extracellular enzymes from Aspergillus cristatus. This study enriched our understanding of Aspergillus cristatus in the aroma formation process of FBT.

Metagenomic insight into the biodegradation of biomass and alkaloids in the aging process of cigar

A significant distinction between cigar production and tobacco lies in the necessary aging process, where intricate microbial growth, metabolic activities, enzymatic catalysis, and chemical reactions interact. Despite its crucial role in determining the final quality of cigars, our comprehension of the underlying chemical and biological mechanisms within this process remains insufficient. Biomass and alkaloids are the primary constituents that influence the flavor of cigars. Consequently, investigating the entire aging process could begin by exploring the involvement of microbes and enzymes in their biodegradation. In this study, handmade cigars were aged under different conditions. Metagenomic sequencing was employed to identify the microbes and enzymes responsible for the degradation of biomass and alkaloids derived from tobacco leaves. The results revealed that various environmental factors, including temperature, humidity, duration time, and turning frequency, yielded varying contents of total sugar and alkaloids in the cigars. Significant correlations were observed between microbial communities and starch, reducing sugars, total sugars, and alkaloids. Key species involved in the breakdown of biomass constituents, such as starch (Bacillus pumilus, Pseudomonas sp. 286, and Aspergillus cristatus), reducing sugars and total sugars (Aspergillus cristatus and Nitrolancea hollandica), were identified. Furthermore, Corynespora cassiicola and Pseudomonas fulva were found to potentially contribute to the degradation of alkaloid compounds, specifically nornicotine and neonicotinoid. Our work contributes to a deeper understanding of the microbial roles in the aging of cigars. Moreover, the selection of specific microbial strains or starter cultures can be employed to control and manipulate the aging process, thereby further refining the flavor development in cigar products.Graphical

Metabolomic and transcriptomic analyses reveal comparisons against liquid-state fermentation of primary dark tea, green tea and white tea by Aspergillus cristatus

Liquid-state fermentation (LSF) of tea leaves is a promising way to obtain tea-based nutraceutical products rich in various bioactive compounds. In the study, the changes of bioactive compounds, tea pigments and complex metabolites from LSF of primary dark tea, green tea and white tea infusions with Aspergillus cristatus were determined. Chemical analyses revealed that soluble sugars, monosaccharide composition, total polyphenols, total flavonoids, free amino acids, soluble proteins and tea pigments were changed in different ways. An untargeted metabolomic analysis and ribonucleic acid sequencing (RNA-seq) based transcriptomic analysis were performed to investigate the metabolic differentiation and clarify the key differentially expressed genes (DEGs, fold change >2 and p < 0.05), showing that amino acid metabolism, carbohydrate metabolism and lipid metabolism were the most enriched pathways during A. cristatus fermentation of primary dark tea, green tea and white tea infusions. In addition, glycerophospholipid metabolism, linoleic acid metabolism and phenylalanine metabolism were greatly accumulated in the fermentation of primary dark tea and white tea infusions; Pyruvate metabolism, glycolysis/gluconeogenesis, fatty acid degradation, tyrosine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis and valine and leucine, isoleucine degradation were greatly accumulated in the fermentation of primary dark tea and green tea infusions; Starch and sucrose metabolism was greatly accumulated in the fermentation of green tea and white tea infusions; Galactose metabolism was significantly enhanced in the fermentation of primary dark tea infusion; Amino sugar and nucleotide sugar metabolism, sphingolipid metabolism and alanine, aspartate and glutamate metabolism were significantly enhanced in the fermentation of green tea infusion. Besides, some other pathways involving aminobenzoate degradation, biosynthesis of cofactors, pyrimidine metabolism, benzoxazinoid biosynthesis and phenazine biosynthesis, tropane, piperidine and pyridine alkaloid biosynthesis and flavone and flavonol biosynthesis also differed from each other. These findings support that A. cristatus plays a vital role in the biochemical and genetic regulation of metabolite profile, and could be considered a potential prospect for better use of A. cristatus on different kinds of tea materials.

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

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

Intracellular Polysaccharides of Aspergillus cristatus from Fuzhuan Brick Tea Leverage the Gut Microbiota and Repair the Intestinal Barrier to Ameliorate DSS-Induced Colitis in Mice.

The intracellular polysaccharides of Aspergillus cristatus (IPSs) from Fuzhuan brick tea have been demonstrated to improve immune function linked to modulating the gut microbiota. Herein, to further investigate the efficacy of IPSs to maintain gut homeostasis, the protection of the purified fraction of IPSs (IPSs-2) on the mice with colitis induced by dextran sulfate sodium (DSS) and the underlying mechanisms were explored in this study. The results revealed that IPSs-2 alleviated the typical symptoms of colitis and suppressed the excessive inflammatory mediators, regulating the genes related to inflammatory responses in the colon at the mRNA level. Meanwhile, IPSs-2 treatment reinforced the intestinal barrier function by ameliorating the DSS-induced histological injury, facilitating the differentiation of goblet cells to enhance Mucin-2 generation, and enhancing the expression of tight junction proteins to alleviate colitis. In addition, IPSs protected against colitis by promoting the production of short-chain fatty acids (SCFAs), the activation of SCFAs receptors, and the leverage of the gut microbiota via the enrichment of Bacteroides, Parabacteroides, Faecalibacterium, Flavonifractor_plautii, and Butyricicoccus, linking with reducing inflammation and repairing intestinal barrier function. Overall, our research revealed the therapeutic potential of IPSs-2 as a prebiotic for attenuating inflammatory bowel disease and provided a rationale for future investigation.

Improved flavonoid content in mulberry leaves by solid-state fermentation: Metabolic profile, activity, and mechanism

This study was aimed at investigating the mechanisms of release and transformation of total flavonoids (TFs) from mulberry leaves (MLs) by solid-state fermentation (SSF) via Aspergillus cristatus. An intelligent model based on a back-propagation artificial neural network (BP–ANN) was established to predict the TFs in fungi co-fermented MLs. After prediction and experimental validation, the TFs released by SSF (72.55 mg rutin equivalents/g dry weight) were significantly higher than that of unfermented (24.42 mg rutin equivalents/g dry weight). In addition, the biotransformation mechanism of flavonoids during SSF was proposed by analyzing the untargeted metabolome, including the flavonoid and phenylpropanoid biosynthetic pathways. Pearson's correlation coefficient analysis indicated that the 29 up-regulated catabolites (especially loureirin D and arachidoside) after SSF contributed to increased ABTS (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) radical scavenging activities and α-glucosidase inhibitory activities. Furthermore, structural characteristics (by scanning electron microscopy, thermogravimetric analysis, and FTIR spectroscopy) and analyses of dynamic changes in carbohydrate-hydrolyzing enzymes revealed that the destruction of hemicellulose was essential to releasing TFs by SFF. Industrial relevanceIn this study, a back-propagation artificial neural network (BP–ANN) was successfully used to establish a co-fermentation with Aspergillus cristatus for releasing and transforming flavonoids of mulberry leaves (MLs) through solid-state fermentation (SSF). The present study has immeasurable importance toward making maximum use of MLs. Moreover, the findings of this study also provide new insights into understanding the release and biotransformation mechanism of flavonoids from MLs by SSF.

Untargeted metabolomics and quantification analysis reveal the shift of chemical constituents between instant dark teas individually liquid-state fermented by Aspergillus cristatus, Aspergillus niger, and Aspergillus tubingensis.

Instant dark teas (IDTs) were individually liquid-state fermented using the fungi Aspergillus cristatus, Aspergillus niger, and Aspergillus tubingensis. To understand how the chemical constituents of IDTs were affected by the fungi, samples were collected and measured by liquid chromatography-tandem mass-tandem mass spectrometry (LC-MS/MS). Untargeted metabolomics analysis revealed that 1,380 chemical constituents were identified in positive and negative ion modes, and 858 kinds of chemical components were differential metabolites. Through cluster analysis, IDTs were different from the blank control, and their chemical constituents mostly included carboxylic acids and their derivatives, flavonoids, organooxygen compounds, and fatty acyls. And the metabolites of IDTs fermented by A. niger and A. tubingensis had a high degree of similarity and were classified into one category, which showed that the fungus used to ferment is critical to the formation of certain qualities of IDTs. The biosynthesis of flavonoids and phenylpropanoid, which involved nine different metabolites such as p-coumarate, p-coumaroyl-CoA, caffeate, ferulate, naringenin, kaempferol, leucocyanidin, cyanidin, and (-)-epicatechin, were significant pathways influencing the quality formation of IDTs. Quantification analysis indicated that the A. tubingensis fermented-IDT had the highest content of theaflavin, theabrownin, and caffeine, while the A. cristatus fermented-IDT had the lowest content of theabrownin, and caffeine. Overall, the results provided new insights into the relationship between the quality formation of IDTs and the microorganisms used in liquid-state fermentation.