Human Fecal Fermentation Research Articles

Structural alterations in butyrylated and recrystallized high-amylose maize starch and their effect on gut microbiota during in vitro fermentation.

In this study, type-3 resistant starch (RS) with enhanced thermal stability and excellent short-chain fatty acid (SCFA) production was obtained through the butyrylation and subsequent recrystallization at 4 °C of high-amylose maize starch (HAMS). We comprehensively examined and contrasted the structural attributes and in vitro human fecal fermentation behavior of butyrylated RS (BRS) with varying degrees of substitution. Fourier-transform infrared analysis validated the successful integration of carbonyl groups into the starch matrix. This phenomenon was evident through the characteristic peaks at 1727 cm-1. X-ray diffraction and thermal stability analyses delineated the distinct B-type crystalline structure and high relative crystallinity of 25.31 % and 22.23 % of the 10 % and 15 % butyric anhydride BRS-10 and BRS-15, respectively, which differed from that of HAMS. Their second peak gelatinization temperature values reached 95.6 °C and 92.6 °C. The in vitro fermentation of BRS fostered SCFA production, boosting the relative abundance of beneficial bacteria (e.g., Ligilactobacillus and Roseburia). Meanwhile, the extensively modified BRS favored the propagation of Faecalibacterium and Bifidobacterium, maintaining intestinal microbiota advantage. These findings underscore the significant effects of butyrylation modification on fermentation kinetics, metabolite profiles, and gut microbiota composition, providing invaluable insights into the development of functional food products that aim to bolster colonic health.

Metabolic outcomes of Cordyceps fungus and Goji plant polysaccharides during in vitro human fecal fermentation

This study was to assess the digestion and colonic fermentation of two bioactive polysaccharides, EPS-LM and LBPS, and the subsequent influences on human gut microbiota through simulated gastrointestinal systems. EPS-LM, an exopolysaccharide isolated from mycelial culture of a medicinal fungus Cordyceps sinensis Cs-HK1, was characterized as a heteropolysaccharide consisting of Man(108):Gal(52.7):Glc(29.2) (molar ratio) with an average molecular weight (MW) 5.513 × 106. LBPS was isolated from a well-known medicinal plant (Lycium barbarum L.) which was also characterized as a heteropolysaccharide (1.236 × 105 MW). Both polysaccharides were highly resistant to saliva, gastric and small-intestine digestion with negligible MW reduction and release of reducing sugars but were quickly degraded to lower MW during in vitro human fecal fermentation. They were consumed as a carbon source by the gut bacteria to produce short-chain fatty acids (SCFAs). In comparison, the carbohydrate content of EPS-LM was more completely consumed than LBPS and there were also notable differences in consumption of specific monosaccharides and production of specific SCFAs, propionic and butyric acid, and relative abundance of gut bacterial populations between EPS-LM and LBPS group. The results suggest that metabolic outcomes and modulating effects of EPS-LM and LBPS on the gut microbiota are highly dependent on their molecular composition.

Comprehensive evaluation of Flammulina velutipes residues polysaccharide based on in vitro digestion and human fecal fermentation

Flammulina velutipes residues (FVR) are the waste culture medium derived from the collection of Flammulina velutipes fruiting bodies, with an annual output that remains largely unexplored. The characteristics of digestion and fermentation of Flammulina velutipes residues polysaccharide (FVRP) are still relatively unknown. This study investigated the structure of the gut microbiota through 16 s rDNA gene sequencing and analyzed changes in short-chain fatty acid (SCFA) content via targeted metabolome analysis. The aim was to explore the prebiotic activity of FVRP based on a simulated digestion model combined with an in vitro anaerobic fermentation model. The results demonstrated that FVRP did not exhibit significant changes during in vitro digestion and fermentation but did enhance antioxidant activity. Furthermore, FVRP was found to rapidly reduce the pH value and increase SCFA production in the fermentation broth from lactic acid bacteria and human feces. Notably, FVRP altered the gut microbiota structure, significantly increasing the relative abundance of Firmicutes and Bacteroidota. Thus, FVRP could be considered a promising prebiotic food and feed additive that promotes the generation of short-chain fatty acids by modulating gut microbiota.

Structural characterization, antioxidant activity, and fermentation characteristics of Flammulina velutipes residue polysaccharide degraded by ultrasonic assisted H2O2-Vc technique

Adhere to the concept of low-carbon environmental protection and turning waste into treasure, polysaccharides from Flammulina velutipes residue polysaccharide (FVRP) has been developed and possesses diverse bioactivities, comprising antioxidant, hypoglycemic, and relieving heavy metal damage, which still has the disadvantages of high molecular weight and low bioavailability. The current work is the first to prepare a degraded polysaccharide (FVRPV) from FVRP by ultrasonic assisted H2O2-Vc technique in order to reduce its molecular weight, thereby improving its activity and bioavailability. Our results found that the molecular weight and average particle size were declined, but the monosaccharide composition and characteristic functional group types of FVRPV had no impact. The structural changes of polysaccharides analyzed by XRD, Congo Red test, I2-KI, SEM, and methylation analysis indicated that the surface morphology and glycosidic bond composition of FVRPV possessed longer side chains and a greater number of branches with an amorphous crystal structure devoid of a triple helix configuration, and had experienced notable alterations after ultrasonic assisted H2O2-Vc treatment. Meanwhile, the in vitro antioxidant capacity of FVRPV had significantly increased compared to FVRP, implying ultrasonic assisted H2O2-Vc technique maybe a effective method to enhance the bioactivity of polysaccharides. In addition, the content of polysaccharide, reducing sugar, and uronic acid in FVRPV was significantly decreased, but antioxidant capacity of fermentation broth was stronger by in vitro human fecal fermentation. The 16S rDNA sequencing data displayed that FVRPV can enrich probiotics and reduce the abundance of pathogenic bacteria through different metabolic pathways mediated by gut microbiota, thereby exerting its potential probiotic effects. The interesting work provides a novel degraded polysaccharide by ultrasonic assisted H2O2-Vc technique, laying a foundation for developing FVRPV as a new antioxidant and prebiotic.

Effects of in vitro simulated digestion and fecal fermentation on the structure and regulating the glucose and lipid activity of a polysaccharide from Mori Folium

Mori folium, as a homologous drug-food, has hypoglycemic and lipid-lowering activity. Polysaccharides are the main bioactive ingredient of the Mori folium that exhibit diverse biological activities. In this study, a homogeneous polysaccharide (MP4) was purified and characterized from Mori folium. The changes of MP4 affected by saliva, simulated gastrointestinal juice, and human fecal fermentation, including physicochemical property or its bioactivity, were systematically investigated. Meanwhile, the influence of fermentation on the bioactivity were evaluated. The results showed that the backbone of MP4 is mainly composed of →4)-α-D-GalpA-(1→ residues. The molecular weight, the levels of reducing sugar content and free monosaccharides of MP4 exhibited no significant differences indicating that gastrointestinal digestion has a minimal effect on the physicochemical characteristics of MP4. However, during in vitro gut microbiota fermentation, MP4 are significantly degraded and utilized by gut microbiota, showing increased the production of short-chain fatty acids, notably acetic acid and propionic acid. The relative abundance of beneficial bacteria such as Bacteroidetes and Actinobacteria were significantly increased, whereas the levels of pathogenic bacteria such as Fusobacteria and Megamonas were significantly decreased, which changed the composition of the gut microbiota. The Firmicutes/Bacteroides ratio was also decreased significantly. Interestingly, after in vitro fermentation, the α-glucosidase inhibitory activity was increased, the lipase inhibitory activity and cholesterol adsorption activity was decreased. Correlation analysis showed that the relative abundance of some bacteria was significantly correlated with the bioactivities. These results provide a basis for the development of Mori folium polysaccharides as functional probiotic products.

Effects of Differently Processed Tea on the Gut Microbiota.

Tea is a highly popular beverage, primarily due to its unique flavor and aroma as well as its perceived health benefits. The impact of tea on the gut microbiome could be an important means by which tea exerts its health benefits since the link between the gut microbiome and health is strong. This review provided a discussion of the bioactive compounds in tea and the human gut microbiome and how the gut microbiome interacts with tea polyphenols. Importantly, studies were compiled on the impact of differently processed tea, which contains different polyphenol profiles, on the gut microbiota from in vivo animal feeding trials, in vitro human fecal fermentation experiments, and in vivo human feeding trials from 2004-2024. The results were discussed in terms of different tea types and how their impacts are related to or different from each other in these three study groups.

Germination-Induced Enhancement of Brown Rice Noodle Nutritional Profile and Gut Microbiota Modulation.

This study explored how germination influences the starch digestion and intestinal fermentation characteristics of brown rice noodle. The study began with in vitro starch digestion tests to assess how germination affects starch digestibility in brown rice noodles, revealing an increase in rapidly digestible starch content and a decrease in resistant starch content. Subsequently, an in vitro human fecal fermentation model was used to simulate the human intestinal environment, showing that germination altered pH levels and the production of short-chain fatty acids, particularly by increasing propionate while decreasing acetate and butyrate. Additionally, the study noted a decrease in gut microbiota diversity following fermentation, accompanied by an increase in Megamonas growth and a decrease in Bacteroides and Bifidobacterium. In conclusion, these findings suggest that germination could enhance the nutritional value and intestinal probiotic properties of brown rice noodles. This research contributes valuable insights into the role of germination in improving the nutritional properties of rice-based products and provides a foundation for further exploration into the development of health-promoting rice noodles.

Simulated human digestion and fermentation of a high-molecular weight polysaccharide from Lentinula edodes mushroom and protective effects on intestinal barrier

Lentinula edodes (Shiitake) is an important edible mushroom and polysaccharides are its major constituents with proven health benefits. The study was to investigate the gut bacterial fermentation and subsequent effects on gut barrier function of a glucan-rich polysaccharide, LePS40 precipitated from the mushroom water extract with 40 % (v/v) ethanol. LePS40 consisted of a β-(1→3)-glucan main chain with substitution in the C-6 position with side chains mainly composed of (1 → 6)-linked β-Glcp residues, (1 → 6)-linked α-Galp residues and terminal residues of β-Glcp. LePS40 was found highly resistant to digestive enzymes and gastric acid in simulated human gastrointestinal tract, but highly fermentable during in vitro human fecal fermentation. The fecal fermentation degradation of LePS40 appeared to selectively break the glucoside linkage in view of the dramatic decrease in the glucose molar ratio (12.68 to 1.07). Compared with the prebiotic reference FOS, LePS40 led to much higher levels of butyric, and propionic acid and a lower level of acetic acid. Moreover, LePS40 enhanced the abundance of some beneficial bacterial populations, but decreased the bacteria possibly linked with fatty-liver disease and colorectal cancer. Furthermore, the fecal fermentation products of LePS40 showed a potential protective effect on intestinal barrier function against inflammatory damage in Caco-2/Raw264.7 co-culture model. These findings suggest the potential of LePS40 for improvement of gut health through modulation of gut microbiota.

Modulation of gut microbiota and metabolites by Flammulina velutipes polysaccharides during in vitro human fecal fermentation: Unveiling Bacteroides as a potential primary degrader

Flammulina velutipes, a widely cultivated species of edible fungus, exhibits diverse functional activities attributed to its polysaccharides. In this study, we employed an in vitro model to investigate the impact of F. velutipes polysaccharides (FVP) fermentation on gut microbiota, with a particular focus on Bacteroides. FVP fermentation resulted in the proliferation of microbiota associated with short-chain fatty acid (SCFA) metabolism and suppression of Escherichia-Shigella. Bacteroides emerged as potential primary degraders of FVP, with species-level analysis identifying the preference of B. thetaiotaomicron and B. intestinalis in FVP degradation. Metabolomics analysis revealed significant increases in hypoxanthine and 7-methyladenine contents, with histidine metabolism emerging as the most enriched pathway. B. nordii and B. xylanisolvens exhibited the most influence on amino acid and SCFA metabolism. Understanding the mechanisms by which gut microbiota metabolize FVP can provide valuable insights into the potential of FVP to promote intestinal health and disease prevention.

Digestive characteristics of Gastrodia elata Blume polysaccharide and related impacts on human gut microbiota in vitro

Ethnopharmacological relevanceGastrodia elata Blume is a traditional Chinese medicine with the effects of improving the deficiency of the body and maintaining health, and polysaccharide (GEP) is one of the effective ingredients to play these activities of G. elata. Traditionally, G. elata is orally administered, so the activities of GEP are associated with digestive and intestinal metabolism. However, the digestive behavior of GEP and its effects on the human gut microbiota are unclear and need to be fully studied. Aim of the studyThis study aimed to investigate the changes in structural characteristics of GEP during digestion and the related impacts of its digestive product on gut microbiota in human fecal fermentation, and to explain the beneficial mechanism of GEP on human health from the perspective of digestive characteristics and “gut” axis. Materials and methodsThe changes of reducing sugars, free monosaccharides and physicochemical properties of GEP during digestion were investigated by GPC, HPLC, FT-IR, CD, NMR, SEM, and TGA. Moreover, polysaccharide consumption, pH value changes, SCFAs production, and changes in gut microbiota during fermentation were also discussed. ResultsDuring digestion of GEP, glucose was partially released causing a decrease in molecular weight, and a change in monosaccharide composition. In addition, the characteristics of GEP before and after digestion, including configuration, morphology, and stability, were different. The digestive product of GEP was polysaccharide (GEP-I), which actively participated in the fecal fermentation process. As the fermentation time increased, the utilization of GEP-I by the microbiota gradually increased. The abundance of probiotics such as Bifidobacterium, Collinsella, Prevotella, and Faecalibacterium was significantly increased, and the abundance of pathogenic Shigella, Dorea, Desulfovibrio, and Blautia was significantly inhibited, thereby suggesting that GEP has the potential to maintain human health through the “gut” axis. In addition, the beneficial health effects of GEP-I have also been observed in the influence of microbial metabolites. During the fermentation of GEP-I, the pH value gradually decreased, and the contents of beneficial metabolites such as acetic acid, propionic acid, and caproic acid significantly increased. ConclusionThe structure of GEP changed significantly during digestion, and its digestive product had the potential to maintain human health by regulating gut microbiota, which may be one of the active mechanisms of GEP.

Probiotic potential of a novel exopolysaccharide produced by Bifidobacterium animalis subsp. Lactis SF

Exopolysaccharides (EPSs) have been shown to possess wide-ranging of physiological functions, including antioxidant, anticancer, and immunomodulatory. However, there is a scarcity of research focused on the probiotic function of EPSs isolated from Bifidobacteria, and further investigation is needed to reveal the mechanisms underlying their functions. In this study, EPSs were isolated and purified from Bifidobacterium animalis subsp. Lactis SF (SF-EPS), and the probiotic potential of SF-EPS were analyzed by in vitro antioxidant capacity assays and human fecal fermentation. Followed by the evaluation of the eps cluster in B. lactis SF. The results showed that SF-EPS (MW: 4.81 × 104 Da) contained glucose, galactose, arabinose, and mannose in a molar ratio of 4.02:1.91:1.02:1.0 and small amounts of glucuronic acid (0.02 ± 0.01 mol%). Furthermore, SF-EPS showed strong antioxidant capacity, which was embodied in its ability of scavenging DPPH and hydroxyl radicals. SF-EPS regulated the gut microbiota by increasing the relative abundances of Faecalibacterium, Anaerostipes, and Bifidobacterium and decreasing the abundance of Enterobacter and Klebsiella. SF-EPS promoted the production of short-chain fatty acids (SCFAs) by intestinal microorganisms. Therefore, SF-EPS has the potential for utilization as a prebiotic in functional foods, enhancing gut health, and offering a natural ingredient for use in industrial food production.

Gut microbiota modulation and effects of a diet enriched in apple pomace on inflammation in a DSS-induced colitis mouse model.

Certain types of soluble dietary fibre, such as pectin and pectic oligosaccharides from different sources, have demonstrated protective effects against inflammation in DSS-induced colitis mouse models. In this work, we have evaluated the impact of a diet enriched in apple pomace (AP-diet), an agricultural by-product with a significant content of pectin and that previously demonstrated prebiotic properties in human fecal batch fermentation models, on the gut microbiota composition, intestinal damage and inflammation markers in a DSS-induced colitis model. We found that the apple pomace enriched diet (AP-diet), providing a significant amount of pectin with demonstrated prebiotic properties, was associated with a slower increase in the disease activity index, translating into better clinical symptomatology of the animals. Histological damage scoring confirmed less severe damage in those animals receiving an AP-diet before and during the DSS administration period. Some serum inflammatory markers, such as TNFα, also demonstrated lower levels in the group receiving the AP-diet, compared to the control diet. AP-diet administration is also associated with the modulation of key taxa in the colonic microbiota of animals, such as some Lachnospiraceae genera and Ruminococcus species, including commensal short chain fatty acid producers that could play a role in attenuating inflammation at the intestinal level.

Tannic acid delaying metabolism of resistant starch by gut microbiota during in vitro human fecal fermentation

This work investigated the effect of tannic acid on the fermentation rate of resistant starch. It was found that 1.0 and 1.5 μmol/L tannic acid decreased the rate of producing gas and short-chain fatty acids (SCFAs) from fermentation of resistant starch, and 1.5 μmol/mL tannic acid had a more profound effect, which confirmed that tannic acid delayed the metabolism of resistant starch. Moreover, tannic acid significantly inhibited the α-amylase activity during fermentation. On the other hand, tannic acid delayed the enrichment of some starch-degrading bacteria. Besides, fermentation of the resistant starch/tannic acid mixtures resulted in more SCFAs, particularly butyrate, and higher abundance of beneficial bacteria, including Bifidobacterium, Faecalibacterium, Blautia and Dorea, than fermentation of resistant starch after 48 h. Thus, it was inferred that tannic acid could delay the metabolism of resistant starch, which was due to its inhibitory effect on the α-amylase activity and regulatory effect on gut microbiota.

Chitobiose exhibited a lipid-lowering effect in ob/ob−/− mice via butyric acid enrolled liver–gut crosstalk

Chitobiose (COS2) efficiently lowers lipids in vivo and facilitates butyric acid enrichment during human fecal fermentation. However, whether COS2 can interact with butyric acid to generate a hypolipidemic effect remains unclear. This study examined the hypolipidemic mechanism of COS2 involving butyric acid, which could alleviate non-alcoholic fatty liver disease (NAFLD). The results revealed that COS2 administration modulated the β-oxidation pathway in the liver and restructured the short chain fatty acids in the fecal of ob/ob−/− mice. Moreover, the hypolipidemic effect of COS2 and its specific accumulated metabolite butyric acid was verified in sodium oleate-induced HepG2 cells. Butyric acid was more effective to reverse lipid accumulation and up-regulate β-oxidation pathway at lower concentrations. Furthermore, structural analysis suggested that butyric acid formed hydrogen bonds with key residues in hydrophilic ligand binding domains (LBDs) of PPARα and activated the transcriptional activity of the receptor. Therefore, the potential mechanism behind the lipid-lowering effect of COS2 in vivo involved restoring hepatic lipid disorders via butyric acid accumulation and liver–gut axis signaling.Graphical

Impact of ultrasound processing of açai juice containing complex carbohydrates in human intestinal microbiota composition and metabolite production

The present study evaluated the impact of high-intensity ultrasound processing of potentially prebiotic açai juice containing complex carbohydrates (dextran and oligosaccharides) on human gut microbiota after in vitro colonic fermentation. The juice was subjected to an in vitro simulated digestion and human fecal fermentation. HPLC quantified the metabolites produced during fecal fermentation, and the 16 S rRNA sequencing evaluated the microbiota change. The gluco-oligosaccharides and dextran were resistant to in vitro digestion and promoted the fecal microbiota production of short-chain fatty acids (SCFAs). The sonicated juice markedly increased propionic and butyric acid production, reaching 11 g/L and 4 g/L, respectively. The potentially prebiotic açai juice improved the richness (25%) and diversity (22%) of the fecal microbiota, and the ultrasound processing increased richness (12%) compared to non-sonicated juice, which may contribute to overall host health. The study demonstrated that açai juice containing gluco-oligosaccharides and dextran has a potential prebiotic effect. Incorporating this juice into the diet might promote gut-beneficial bacterial growth, improving the gut microbiota and the health-promoting metabolites.

Antioxidant Activities and Prebiotic Activities of Water-Soluble, Alkali-Soluble Polysaccharides Extracted from the Fruiting Bodies of the Fungus Hericium erinaceus.

In this study, the digestion and fermentation properties of the bioactive water-soluble polysaccharide (HEP-W), and alkali-soluble polysaccharide (HEP-A) from Hericium erinaceus and the impact on the human colonic microbiota were determined using simulated saliva-gastrointestinal digestion and human fecal fermentation models in vitro. The basic physicochemical properties of HEP-W and HEP-A were determined at the same time. The results showed that the in vitro simulated digestion had almost no effect on the physicochemical properties of HEP-W and HEP-A, indicating that HEP-W and HEP-A were partially degraded. During fermentation, HEP-W and HEP-A increased the relative abundance of the dominant butyric acid-producing genera, the microbial community structure was significantly regulated, the gas production and short-chain fatty acid production in the fermentation broth were significantly increased, and the pH of the fermentation broth was reduced. There were structural and other differences in HEP-W and HEP-A due to different extraction methods, which resulted in different results. These results suggest that HEP-W and HEP-A may be potential gut microbial manipulators to promote gut health by promoting the production of beneficial metabolites by intestinal microorganisms using different butyric acid production pathways.

Modulating in vitro fecal fermentation behavior of sodium alginate by Ca2+ cross-linking

Slow fermentable dietary fibers can be utilized by human gut microbiota in the distal region of the colon and thus exert a sufficient short-chain fatty acids (SCFAs) supplement in the distal region of the human colon. Alginate (Alg) based microgels are widely fabricated and used to control their digestion by digestive enzymes releasing active substances site-specifically. Herein, sodium alginate microgels with gradient calcium-ion (Ca2+) cross-linking densities were developed, restricting their degradation by gut microbiota. Alg microgels were prepared using high-speed shearing after Alg was cross-linked with 10, 40, and 60 mmol/L Ca2+, respectively (named 10-Alg, 40-Alg, and 60-Alg). The fluorescence and atomic force microscopic results showed that the 40-Alg particle has the densest structure among the three cross-linked Alg. In vitro human fecal fermentation results revealed that the Ca2+ cross-linking exerted more restricting effects than delaying effects on the fermentation of Alg, and the 40-Alg exhibited the slowest fermentation rate and the least fermentation extent, by characterizing the residual total carbohydrate content, residual monosaccharide content, pH, and total short-chain fatty acids. The 16S rRNA gene sequencing results indicated that cross-linking structures shaped a high specifical Bacteroides-type microbial community and that OTU205 (Bacteroides_xylanisolvens) highly correlated to the cross-linking density (R = 0.65, p = 0.047). In sum, Ca2+ cross-linking generated a dense and compact structure of sodium alginate that facilitated a more restricted fermentation property and specificity-targeting microbial community structure in comparison to the original sodium alginate.

Enzymatic production of cello-oligosaccharides with potential human prebiotic activity and release of polyphenols from grape marc

Grape marc, the main winemaking byproduct, is an excellent source of bioactive polyphenols, such as anthocyanins, resveratrol and quercetin. An enzyme-catalysed treatment of marc was developed using endo-1,4-β-d-glucanase to release polyphenol O-glucosides and simultaneously generate the optimal concentration of water-soluble cello-oligosaccharides (COS), including cellopentaose, cellotriose, and cellobiose from the marc matrix. The prebiotic properties of marc hydrolysate rich in COS was assessed using human probiotic monocultures of Lactobacillus spp. and Bifidobacterium spp. strains, and invitro human faecal fermentation. The COS-rich hydrolysate showed excellent prebiotic effect in both studies, successfully supporting the growth of beneficial probiotic strains, and was highly fermentable by faecal microbiota producing gas and short chain fatty acids. Acetate and propionate production were the highest when faecal bacteria fermented COS-rich solution compared with standard substrates. For the first time, COS was shown to be fermented by faecal microbiota, demonstrating the potential benefits of valorised grape marc.

Fecal fermentation and gut microbiota modulation of dietary fibre and polyphenols from Gnetum gnemon Linn. leaves

The purpose of this study was to analyze the effects of Gnetum gnemon var.tenerum leaf powder, dried using a vacuum microwave, on gut health properties. The two alternative drying conditions were 3600 Watts for 12 min (G1) and 2400 Watts for 15 min (G2) compared to the commercial Japanese green tea matcha powder (M) and the gut health effects were assessed by human fecal fermentation in a one-stage colon system. The enumeration of gut microbiota at 24 h of fermentation by next generation sequencing (NGS) confirmed elevated Bacteroides levels and significantly (P < 0.05) higher numbers of Bifidobacterium with G1 (33%) and G2 (37%) compared to M (31%). Additionally, short-chain fatty acid (SCFA) profile for acetic, propionic and butyric acids was analysed using gas chromatography-flame ionization detector (GC-FID), and especially butyric acids was significantly (P < 0.05) higher with G1 (4.698 ± 0.61 mM) and G2 (2.768 ± 0.10 mM) compared to M, 1.805 ± 0.01 mM at 24 h. The Gnetum leaf powders G1 and G2 were investigated for promoting γ-aminobutyric acid (GABA) microbial metabolite, which was detected using liquid chromatography-mass/mass spectrometry (LC-MS/MS). Thus, Gnetum gnemon var. tenerum leaf powder could provide gut health benefits with good potential for use as a functional food ingredient in the future.

Evaluation of the Enzymatic Production and Prebiotic Activity of Galactomannan Oligosaccharides Derived from Gleditsia microphylla

Oligosaccharides have received considerable attention as prebiotics because they exhibit potential health benefits related to their ability to modulate intestinal bacterial composition. This study evaluated the effects of galactomannan oligosaccharides (GMOS) derived from Gleditsia microphylla as a prebiotic on human intestinal bacteria. The β-mannanase used for the enzymatic hydrolysis of GMOS was produced by Trichoderma reesei Rut C-30. The enzymatic hydrolysis of GMOS was found to occur under optimal conditions at 50 °C, pH 5, 20 U/g-GM, and 20 g/L, and resulted in a yield of 70.78% ± 1.34%. The purity of GMOS after purification was 81.50%. Upon performing in vitro human fecal fermentation using GMOS as a carbon source, it was observed that GMOS effectively promoted the proliferation of intestinal bacteria, and the utilization efficiency of GMOS by intestinal bacteria was found to be at 98.40%. In addition, GMOS were found to have a stabilizing effect on intestinal pH. Additionally, 16S rRNA sequencing of GMOS revealed that GMOS significantly affected the diversity of gut microbiota. Specifically, GMOS exhibited a significant inhibitory effect on Fusobacteria at the phyla and genus level, and demonstrated a significant inhibitory effect on Fusobacterium. Moreover, the results for the prediction of metabolic function analysis showed that GMOS had a significant effect on the level two metabolism of carbohydrates, cofactors, and vitamins. Furthermore, during level three metabolism, the lipoic acid metabolism was significantly affected by GMOS. These results provide a theoretical basis for the potential use of galactomannan oligosaccharides from Gleditsia microphylla as prebiotics for regulating human intestinal bacteria.