Metabolic and microbial modulation of phenolic compounds from raspberry leaf extract under in vitro digestion and fermentation

Raspberry leaves, by-products in raspberry production, are also a rich source of bioactive phytochemicals. In this study, the changes of phenolic compounds in raspberries leaf extract (RLE) under in vitro digestion and colonic fermentation were further studied by HPLC-MS analysis and 16S rRNA. The results showed that the phenolic compounds in RLE were relatively stable during in vitro gastric digestion; however, in the subsequent intestinal digestion and colonic fermentation, their content decreased sharply. A large amount of hydroxyphenylpropionic acid, hydroxyphenylacetic acid and urolithins were produced under the action of gut microbiota. Moreover, compared with corresponding control, RLE significantly reduced the ratio of Firmicutes/Bacteroidetes in all volunteers, increased the relative abundances of some beneficial bacteria, Enterococcus, Prevotella, and decreased the relative abundances of potential pathogens, Clostridium and Faecalibacterium. These findings suggest that RLE during in vitro digestion and fermentation has positive effects on gut microbiota and potential value of maintaining intestinal health.     

Changes in polyphenol fractions and bacterial composition after in vitrofermentation of apple peel polyphenol by gut microbiota

The changes in polyphenol fractions after in vitro fermentation of apple peel polyphenol (APP) by gut microbiota as well as the effects of APP on the growth, pH value, short-chain fatty acids (SCFAs) production and intestinal flora composition of gut microbiota fermentation were firstly explored in this study. The relative abundance of Lactobacillus in the APP group was 49.55% with an increment of 49.40% compared with the blank group, while that of Bifidobacterium was 13.32%. Moreover, the flora produced 37.093 ± 0.478 mM of SCFAs including acetic acid, propionic acid, butyric acid and valeric acid during the process of fermentation, thus reduced the environmental pH value. In addition, polyphenol fractions in APP were altered by gut microbiota fermentation to some extent, for example, glycosides were hydrolysed to aglycones. These findings suggested APP as a potential prebiotic agent to alleviate the disorder of intestinal flora.     

In vitro caecum fermentation and in vivo (Gallus gallus) of calcium delivery systems fabricated by desalted duck egg white peptides and chitosan oligosaccharide on gut health

As calcium supplements, two novel calcium delivery systems were prepared with desalted duck egg white peptides (DPs) and chitosan oligosaccharide (COS) by Amadori-linkage and transglutaminase (TGase)-induced reaction, with high calcium-binding ability of promoting calcium absorption in vivo and in vitro. However, there are relatively few studies on the gut health of calcium delivery systems. In this experiment, in vitro caecum fermentation experiment and Gallus gallus intra amniotic feeding model were used to assess the gut health of calcium delivery systems and phytic acid. Results showed that two calcium delivery systems improved the microbial community structure by inhibiting the proliferation of Escherichia coli, and promoting the proliferation of Bifidobacterium and Lactobacillus. The addition of calcium delivery systems significantly increased short chain fatty acids (SCFAs) and improved the boundary membrane function of brush nutrients by significantly increasing villi structure, surface area, cup cell expansion and production of associated genes (P < 0.05). Moreover, calcium delivery systems present significant reverse of the inhibitory effect of phytic acid on gut health through in vitro and in vivo experiments. This study suggested that DPs-COS-Ca systems were superior to the commercially available calcium supplement CaCO₃ and had excellent calcium absorption and gut health regulating effects.     

枸杞多糖体外调节人体肠道菌群的功能研究

为研究枸杞多糖(LBP)对人体肠道菌群结构和代谢产物的影响,对6名健康人的粪便提取物进行单独样本与混合样本的体外模拟厌氧发酵,利用16S rRNA基因高通量测序技术对发酵后肠道菌群进行结构和功能分析,并利用超高效液相色谱(UPLC)检测发酵液中的短链脂肪酸(SCFAs)浓度。结果表明,LBP能够明显改变人体肠道菌群结构与功能,提高肠道菌群中益生菌乳酸杆菌属与双歧杆菌属的丰度,并促进了SCFAs的产生。因此,LBP能够显著影响人体肠道菌群结构与功能。     

Influence of Intestinal Microbiota on the Catabolism of Flavonoids in Mice

Although in vitro studies have shown that flavonoids are metabolized into phenolic acids by the gut microbiota, the biotransformation of flavonoids by intestinal microbiota is seldom studied in vivo. In this study, we investigated the impact of the gut microbiota on the biotransformation of 3 subclasses of flavonoids (flavonols, flavones, and flavanones). The ability of intestinal microbiota to convert flavonoids was confirmed with an in vitro fermentation model using mouse gut microflora. Simultaneously, purified flavonoids were administered to control and antibiotic‐treated mice by gavage, and the metabolism of these flavonoids was evaluated. p‐Hydroxyphenylacetic acid, protocatechuic acid, p‐hydroxybenzoic acid, vanillic acid, hydrocaffeic acid, coumaric acid, and 3‐(4‐hydroxyphenyl)propionic acid were detected in the serum samples from the control mice after flavonoid consumption. The serum flavonoid concentrations were similar in both groups, whereas the phenolic metabolite concentrations were lower in the antibiotic‐treated mice than in the control mice. We detected markedly higher flavonoids excretion in the feces and urine of the antibiotic‐treated mice compared to the controls. Moreover, phenolic metabolites were upregulated in the control mice. These results suggest that the intestinal microbiota are not necessary for the absorption of flavonoids, but are required for their transformation.     

Assessment of the probiotic potential of lactic acid bacteria isolated from Minas artisanal cheese produced in the Campo das Vertentes region,Brazil

The microbiota of the Brazilian Minas artisanal cheese, made from raw milk, is not well known and may include probiotic bacteria. This study aimed to assess the in vitro and in vivo probiotic properties of the lactic acid bacteria isolated from these cheeses. Thirty‐six samples of the Lactobacillus/Pediococcus group were selected for in vitro investigation. Pediococcus acidilactici (PA2) and Lactobacillus plantarum (LP4) showed the best results and were tested for their ability to protect Salmonella Typhimurium orally infected mice. LP4 showed better probiotic potential than PA2 and allowed higher values of weight gain (P < 0.05). Thus, Lb. plantarumLP4 may have potential use as a probiotic in foods after future technological screening.     

Perspectives of lupine wholemeal protein and protein isolates biodegradation

Lupine (Lupinus angustifolius L.) protein (in wholemeal and protein isolates) was biodegraded using Pediococcus acidilactici in submerged and solid-state fermentation conditions. The changes in the molecular weight of lupine protein fractions, amino acid (AA) profile, biogenic amine formation, antimicrobial and antioxidant properties, and protein digestibility in vitro and in vivo (in Wistar rats) were evaluated. After biotreatment, lower molecular weight peptides (from 10 to 20 kDa) were established, and the free AA content increased. Biodegradation improved the antioxidant properties, modulated the antimicrobial properties, and led to higher in vitro and in vivo digestibility and functionality of the lupine in treated rats (significant increase in body weight of Wistar rats, and increased acetic acid concentration and lowered Escherichia coli count in the caecum). Overall, the biodegradation of lupine protein can transform the plant protein, producing enhanced functionality and bioavailable products.     

Simulated gastrointestinal digestion and in vitro colonic fermentation of date (Phoenix dactylifera L.) seed polyphenols

Date seeds are a by‐product of date fruit industry and a rich source of polyphenols. In this study, in vitro bioaccessibility and colonic fermentation of major polyphenols from date seed powder (DSP) and DSP‐fortified yoghurt (DSPY) were investigated using HPLC. Catechin, epicatechin and procyanidin A2, B1 and B2 were stable during simulated gastric and sequential intestinal digestion. Bioaccessibility was significantly (P < 0.05) higher for all compounds from DSPY compared with DSP. After in vitro colonic fermentation of insoluble digestion materials, most of the target compounds were metabolised by faecal bacteria to ferulic acid, 3‐hydroxyphenylacetic acid, 3‐phenylpropionic acid and 3‐(4‐hydroxyphenyl) propionic acid. DSPY contained significantly (P < 0.05) higher level of free polyphenols as indicated by higher bioaccessibility; however, the stability of the polyphenols and their fermentation products from DSPY were similar to that of DSP alone. These data would be useful in product developments incorporating DSP as a source of polyphenols in food products.     

Partial replacement of meat by sugar cane fibre: cooking characteristics,sensory properties of beef burgers and in vitro fermentation of sugar cane fibre

In this study, sugar cane fibre (SCF) partially replaced meat in beef burger formulations. The effects of SCF on cook yield, dimensional changes, sensory characteristics of beef burgers and in vitro gut fermentation characteristics were evaluated. Replacing beef with 1 to 5% SCF in burgers significantly increased cook yields from 13.8 ± 0.3 to 59.1 ± 0.3% due to its high water-binding capacity of 5.89 ± 0.08 g g⁻¹ and oil-binding capacity of 4.68 ± 0.03 g g⁻¹. The inclusion of SCF improved cooking properties whilst improving sensory characteristics. Burgers with 1% SCF had the highest overall acceptability. SCF was steadily fermented with a porcine faecal inoculum for up to 72 h, producing short-chain fatty acids. The characteristics of high water/oil binding and fermentability suggest that SCF has the potential to provide a range of dietary fibre benefits, and therefore deserves further study.     

Evaluation of the prebiotic potential of five kiwifruit cultivars after simulated gastrointestinal digestion and fermentation with human faecal bacteria

We studied the in vitro prebiotic potential of five different cultivars of kiwifruit including the green‐fleshed ‘Hayward’ and ‘Zesh004’ and the gold‐fleshed ‘Hort16A’, ‘Zesy002’ and ‘Zesy003’. The kiwifruit (25 g fresh weight equivalent) were subjected to simulated gastrointestinal digestion before fermentation for 16 h with faecal microbiota from ten individual donors. Microbial metabolites including lactate were quantified while changes in microbiome composition were determined by 16S rRNA sequencing. Lactate concentrations were highest with ‘Hayward’ (P = 0.01) and correlated with the amount of the kiwifruit fibre and polyphenols, chlorogenic acid and cryptochlorogenic acid. All the kiwifruit behaved similar to inulin in increasing the relative abundance of Bifidobacterium (P < 0.001), but unlike inulin, significantly (P < 0.001) increased the abundances of Ruminococcaceae and decreased Bacteroides. In comparison with inulin, the green‐fleshed kiwifruit selectively increased Lachnospira (P = 0.008) while the gold‐fleshed kiwi fruit increased Akkermansia (P < 0.001). These data suggest that the fibre and polyphenol content of the kiwifruit play a role in modulating gut microbial metabolism. Further clinical studies with these kiwifruit cultivars are required to confirm the potential prebiotic benefits that may be achieved by normal dietary intervention.     

海藻糖基麦芽五糖及其微球的体外消化和酵解

为研究包埋前后的海藻糖基麦芽五糖(N-G7)在模拟消化液及体外酵解过程中的变化,采用口腔、胃部及小肠的体外消化模型分析消化前后溶液中N-G7的质量和N-G7微球(ALGCa)中的糖组成变化,应用体外发酵技术评价N-G7微球的发酵行为,检测不同时间点的pH值、菌体浓度(OD₆₀₀)、产气量、短链脂肪酸(SCFA)浓度等指标的变化,并探究其对人体肠道微生物构成的影响。结果表明：N-G7在口腔消化2 min后在强酸性的胃部环境中几乎不被水解,在小肠中N-G7的水解率达到了82.70%。采用不同质量分数(1%、2%、3%)的海藻酸钙对其进行包埋,形成的微球保留80%以上的N-G7到达结肠。在体外酵解中,各实验组的pH随时间延长逐渐降低,OD600随时间延长逐渐增加。质量分数2%的ALG-Ca组产气量和碳源消耗率在0～6 h内达到最低,而后逐渐增加,证明包埋后的N-G7发酵体系属于缓慢发酵。与对照相比,质量分数2%的ALG-Ca组产生大量SCFA,肠道菌群组成发生变化,更有利于益生菌的生长。     

Influence of physiological and chemical factors on the absorption of bioactive peptides

Food-derived bioactive peptides can be released during digestion and have a potentially beneficial effect on human health. It is known that the penetration of peptides across the small intestine can take place in several ways. The research related to the bioavailability of peptides develops dynamically, but still raises many doubts. Since there are many barriers to absorption of peptides, there is still a need to carry out in vitro and in vivo investigations on the bioavailability of these compounds. Moreover, there are few in vivo studies regarding the absorption and activity of peptides in organs, hence the lack of unequivocal and reliable information about this process. Therefore, this review discusses the current research on in vitro and in vivo absorption of these compounds and factors that play a key role in this process and provides better understanding of the in vitro and in vivo bioavailability and bioaccessibility of peptides.     

Optimisation of sourdough bread incorporation into wheat bread by response surface methodology: Bioactive and nutritional properties

The optimisation of the quantity of sourdough (A) prepared with two different fermentation methods, [spontaneous fermentation (F1) vs. starter of lactic acid bacteria‐added fermentation (F2)], instant active dry yeast (B) and wheat bran (C) for a nutritionally improved bread formulation has been studied by evaluating the bioavailability and bioactive properties. The bread produced according to the optimised formula and fermentation types of F1 (OB_F1) and F2 (OB_F2) was compared with control bread (CB). The optimised levels for F1 were 11.45% for sourdough, 1.10% for dry yeast and 1.58% for wheat bran and for F2 6.99% for sourdough, 1.02% for dry yeast and 38.84% for wheat bran. The addition of sourdough significantly (P < 0.05) affected antioxidant activity, total phenolic content, in vitro ash and protein digestibility, and enzyme resistance starch contents of bread. The F1 fermentation method was found to be more effective in terms of bread properties examined.     

Different Flavonoids Can Shape Unique Gut Microbiota Profile In Vitro

The impact of flavonoids has been discussed on the relative viability of bacterial groups in human microbiota. This study was aimed to compare the modulation of various flavonoids, including quercetin, catechin and puerarin, on gut microbiota culture in vitro, and analyze the interactions between bacterial species using fructo‐oligosaccharide (FOS) as carbon source under the stress of flavonoids. Three plant flavonoids, quercetin, catechin, and puerarin, were added into multispecies culture to ferment for 24 h, respectively. The bacterial 16S rDNA amplicons were sequenced, and the composition of microbiota community was analyzed. The results revealed that the tested flavonoids, quercetin, catechin, and puerarin, presented different activities of regulating gut microbiota; flavonoid aglycones, but not glycosides, may inhibit growth of certain species. Quercetin and catechin shaped unique biological webs. Bifidobacterium spp. was the center of the biological web constructed in this study.     

Physicochemical properties, α-amylase and α-glucosidase inhibitory effects of the polysaccharide from leaves of Morus alba L. under simulated gastro-intestinal digestion and its fermentation capability in vitro by human gut microbiota

The investigation aimed at determining the impact of sequential simulated digestion on the physicochemical properties and digestive enzymes inhibitory effects of the polysaccharides fraction (MLP-2) of Morus alba L. leaves as well as its in vitro fermentation behaviours. After artificial salivary, gastric and intestinal digestions, the chemical components and microstructure of MLP-2 were altered with significantly (P < 0.05) decreased molecular weight. The α-amylase and α-glucosidase inhibitory activities of MLP-2 were significantly (P < 0.05) improved throughout simulated digestion. MLP-2I, the intestinal digested fraction of MLP-2, could significantly (P < 0.05) decrease the pH value of fermented culture and increase the short-chain fatty acids (SCFA) concentrations, especially acetic, propionic and butyric acids. In conclusion, MLP-2 could be gradually degraded under simulated digestion with altered physicochemical properties and enhanced α-amylase and α-glucosidase inhibitory effects, and further utilised by human gut microbiota to decrease pH value and promote SCFA production.     

Dietary Casein and Soy Protein Isolate Modulate the Effects of Raffinose and Fructooligosaccharides on the Composition and Fermentation of Gut Microbiota in Rats

Although diet has an important influence on the composition of gut microbiota, the impact of dietary protein sources has only been studied to a minor extent. In this study, we examined the influence of different dietary protein sources regarding the effects of prebiotic oligosaccharides on the composition and metabolic activity of gut microbiota. Thirty female rats were fed casein and soy protein isolate with cellulose, raffinose (RAF), and fructooligosaccharides (FOS). Microbiota composition was examined by real‐time qPCR and denaturing gradient gel electrophoresis. Dietary protein source affected cecum microbiota; acetic acid concentration and Lactobacillus spp. populations were greater with soy protein than with casein. Prebiotic oligosaccharides had distinctive effects on gut microbiota; RAF increased the acetic acid concentration and Bifidobacterium spp. populations, and FOS increased the butyric acid concentration regardless of the dietary protein. Likewise, Bifidobacterium sp., Collinsella sp., and Lactobacillus sp. were detected in microbiota of the rats fed RAF, and Bacteroides sp., Roseburia sp., and Blautia sp. were seen in microbiota of the rats fed FOS. Interactions between dietary proteins and prebiotic oligosaccharides were observed with Clostridium perfringens group populations and cecum IgA concentration. RAF and FOS decreased C. perfringens group populations in casein‐fed rats, and the combination of soy protein and RAF substantially increased cecum IgA concentration. These results indicate that dietary proteins can differentially modulate the effects of prebiotic oligosaccharides on gut fermentation and microbiota, depending on the type of carbohydrate polymers involved.     

Current and future experimental approaches in the study of grape and wine polyphenols interacting gut microbiota

Interactions between polyphenols and gut microbiota are indeed a major issue of current interest in food science research. Knowledge in this subject is progressing as the experimental procedures and analysis techniques do. The aim of this article is to critically review the more leading-edge approaches that have been applied so far in the study of the interactions between grape/wine polyphenols and gut microbiota. This is the case of in vitro dynamic gastrointestinal simulation models that try to mitigate the limitations of simple static models (batch culture fermentations). More complex approaches include the experimentation with animals (mice, rats, pigs, lambs and chicks) and nutritional intervention studies in humans. Main advantages and limitations as well as the most relevant findings achieved by each approach in the study of how grape/wine polyphenols can modulate the composition and/or functionality of gut microbiota, are detailed. Also, common findings obtained by the three approaches (in vitro, animal models and human nutritional interventions) such as the fact that the Firmicutes/Bacteroidetes ratio tends to decrease after the feed/intake/consumption of grape/wine polyphenols are highlighted. Additionally, a nematode (Caenorhabditis elegans) model, previously used for investigating the mechanisms of processes such as aging, neurodegeneration, oxidative stress and inflammation, is presented as an emerging approach for the study of polyphenols interacting gut microbiota. © 2020 Society of Chemical Industry     

Effects of particle size on physiochemical and in vitro digestion properties of durum wheat bran

In order to promote the potential health benefits of high fibre products, wheat bran is the main focus of the food industry. The physiochemical and in vitro digestion properties of wheat bran containing different particle size were investigated and compared against raw bran samples. Firstly, the bran sample containing superfine particles (11.63 μm) was hydrolyzed by the α-amylase, pepsin, and pancreatin enzymes separately using a single factor and orthogonal test. Secondly, optimized hydrolysis parameters of superfine particles were employed to measure in vitro digestibility of macronutrients in raw, coarse, medium, and superfine bran particles. The maximum degree of hydrolysis obtained via α-Amylase (concentration 10 mg mL⁻¹, pH 6.6, and time 12.5 min) was 55.71%; pepsin (concentration 50 mg mL⁻¹, pH 1.2, and time 9 h) was 82.10%; and pancreatin (concentration 100 mg mL⁻¹, pH 7.0 and time 12 h) was 84.71%, respectively. The highest in vitro digestibility rate of reducing sugar, protein, fat, and soluble fibre content was observed in superfine bran samples to 33.4%, 82.55%, 91.53%, and 21.66%, respectively.     

Impact of whey protein addition on wheat bread fermented with a spontaneous sourdough

Wheat bread is consumed in large quantities throughout the world. However, it contains an inadequate amount of proteins. This fact can be improved by adding whey protein concentrate (WPC). Sourdough (S) fermentation generally improves some characteristics and the digestibility of bread. The aim of this paper was to determinate the physicochemical properties and in vitro protein digestibility of bread with 20% of WPC leavened with spontaneous S (WPC 20-S). To complete the experimental design, bread with S without WPC (WPC 0-S) and bread with yeast (Y) without and with WPC 20% (WPC 0-Y and WPC 20-Y, respectively) were produced. WPC 20-S bread had higher pH, lower moisture and higher hardness and chewiness, compared with WPC 0-S. Interestingly, WPC had a negative effect only on the specific volume of bread fermented with Y but not when S was used. For all formulations, WPC improved the protein content and in vitro protein digestibility.