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The objective of this study was to identify bioactive oligosaccharides and peptides in the cooking water of chickpeas and common beans, known as aquafaba. The oligosaccharides stachyose, raffinose and verbascose were quantified by high-performance anion-exchange chromatography; 78 and 67 additional oligosaccharides were identified in chickpea and common bean aquafaba, respectively, by LC-MS/MS. Chickpea aquafaba uniquely harbored ciceritol and other methyl-inositol-containing oligosaccharides. In prebiotic growth assays, chickpea aquafaba oligosaccharides were differentially utilized, promoting growth of Limosilactobacillus reuteri DSM 20016 and Bifidobacterium longum subsp. infantis ATCC 15697, but not Lacticaseibacillus rhamnosus GG. Dimethyl labeling, along with LC-MS/MS, effectively differentiated α- and γ-glutamyl peptides, revealing the presence of several γ-glutamyl peptides known to possess kokumi and anti-inflammatory activities, including γ-Glu-Phe and γ-Glu-Tyr in chickpeas aquafaba and γ-Glu- S -methyl-Cys and γ-Glu-Leu in beans aquafaba. This work uncovered unique bioactive peptides and oligosaccharides in aquafaba, helping promote its valorization, food system sustainability, and future health-promoting claims. • 81 and 70 oligosaccharides were identified in chickpea and bean aquafaba by LC-MS/MS. • Chickpea aquafaba oligosaccharides were differentially used by probiotic bacteria. • 332 and 350 peptides were identified in chickpea and bean aquafaba by LC-MS/MS. • Dimethyl labeling and LC-MS/MS revealed the presence of several γ-glutamyl peptides. • Uncovering bioactive compounds in aquafaba helps promote its valorization. [ABSTRACT FROM AUTHOR]

Highlights • Transformation of soy milk isoflavones by bifidobacteria strains was investigated. • Both Bifidobacterium species were able to produce equol. • Optimization of equol production was carried out. • Role of pH, temperature and inulin on equol production in soymilk by bifidobacteria. • Results found the highest equol production was at pH 8, 30 °C and 0.5% inulin. Abstract This study analyzed the effect of pH (X1), temperature (X2) and inulin amount (X3) on transformation of isoflavones (daidzin and daidzein) to equol in soymilk fermented with Bifidobacterium spp. All responses significantly (p < 0.05) fitted into quadratic models with coefficients of determination (R2) close to 1 (0.935–0.989). At 24 h of fermentation, amounts of daidzin and daidzein were influenced by all factors. While at 48 h, all factors affected daidzin and only temperature affected daidzein. Equol production was influenced by pH and temperature in 24 h and by all factors in 48 h fermentation. The optimum conditions for equol production were pH 8, 30 °C and 0.5% inulin. Model validation demonstrated there was no significant (p > 0.05) difference between the experimental and predicted values, suggested the suitability of established models in explaining the daidzin and daidzein transformation to equol as a function of pH, temperature and inulin. [ABSTRACT FROM AUTHOR]

Highlights • B. longum was encapsulated in soy protein isolate-carrageenan complex coacervates. • Complex coacervates produced at pH 3 had higher yields and entrapment efficiency. • Best viability was achieved for 10:1 soy protein isolate-carrageenan ratio. Abstract Soy protein isolate (SPI) and carrageenan (IC) complex coacervates were formed through electrostatic attractions for encapsulating Bifidobacterium longum. The effects of pH (2.0–5.0) and SPI:IC mass ratios (10:1, 15:1, 20:1) on coacervate yield, entrapment efficiency and viability of the probiotic bacteria were investigated. The coacervates produced at pH 3 had higher yields and entrapment efficiency, and a SPI:IC mass ratio of 10:1 produced a complex coacervate with more compact microstructure. Compared to the native ones, the bacteria encapsulated in the coacervates had significantly improved viability during storage (4 °C), pasteurization (85 °C for 5, 10 and 30 min) and in vitro dynamic gastric and intestinal digestion. The findings also suggested that the coacervate with a SPI:IC ratio of 10:1 was more capable to protect the bacteria from loss against different stresses. This study provides a novel approach for designing efficient microcapsules containing probiotic bacteria with enhanced functional properties. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the proliferation effect of fractionated lotus seed resistant starch (LRS3-20%, LRS3-30%) and the structural changes following fermentation. Fractionated lotus seed resistant starch, especially LRS3-20%, promoted the proliferation of these probiotics, and Bifidobacterium longum produced large quantities of acetic acid, while Lactobacillus delbrueckii subsp. bulgaricus produced lactic acid. Additionally, the probiotics with lotus seed resistant starch (LRS3) and LRS3-20% survived more successfully in simulated gastrointestinal solutions compared to glucose, LRS3-30% and high amylose maize starch, but LRS3 and LRS3-20% differed in their abilities to enhance the tolerance to acidic buffer, bile acid and simulated gastric juice. Furthermore, after fermentation, fractionated lotus seed resistant starch displayed a loose, rough surface. The crystalline and double helical structure increased as the amount of amorphous and sub-crystalline structure reduced following fermentation. Interestingly, structural changes were correlated with the original structures and utilisation by the probiotics. [ABSTRACT FROM AUTHOR]

[Display omitted] • Gelatin/gellan gum bilayer film was prepared as a novel probiotic carrier material. • Blueberry anthocyanin extract (BE) improved the probiotic viability in the film. • Baobab seed oil (BO) improved water and oxygen barrier properties of the stored film. • BO/BE loaded-bilayer film obtained the greatest Bifidobacterium longum viability. • BO/BE loaded-bilayer film maintain satisfactory B. longum viability in steamed bread. The gelatin/gellan gum based-bilayer emulsion film was developed in this work to improve the survivability of Bifidobacterium longum during the storage process. The baobab seed oil (BO) was added to the gelatin (GE) matrix to develop emulsion film as the barrier outer layer. The blueberry anthocyanin extract (BE) was incorporated into the gellan gum (GG)-based inner layer to enhance the viability of B. longum. The SEM and FTIR results revealed that the probiotics were successfully entrapped in BO/BE-loaded bilayer film. The greatest survivability and viable cell numbers of the B. longum during the storage period were observed in the BO/BE loaded bilayer film. Furthermore, the stability of the colorful patterns by electrochemical writing was also evaluated in this work. Finally, the GE/BO-GG/BE/BM maintain satisfactory probiotic viability in steamed bread coating application. Hence, the GE/BO-GG/BE/BM bilayer film could be considered a novel material to deliver and protect the probiotics in food applications. [ABSTRACT FROM AUTHOR]

[Display omitted] • Lotus seed oligosaccharides (LOS) monomer enriched beneficial bacteria in vivo. • The responses of Bifidobacterium species to LOS were studied in vitro. • The glycolysis site metabolism of LOS4 by B. adolescentis were obtained. • The structure-prebiotic function relationship of LOS was obtained. • Probiotics showed a higher structural preference. Natural lotus seed oligosaccharides monomers (LOSs: LOS3-1, LOS3-2, and LOS4) were prepared by preparative chromatography and were hydroxyl-labeled with fluorescein isothiocyanate (FITC). The prebiotic properties of LOSs by the gut microbiota of male Balb/C mice in vivo and in vitro were studied. In vivo experiment results showed that LOS4 could significantly increase the average daily food consumption, weight, liver index and the abundance of Bacteroides and Bifidobacterium for mice (p < 0.05). In addition, LOS4 also had significant proliferation effect on Bifidobacterium adolescentis and longum in vitro (p < 0.05). Laser confocal microscopy observation showed interaction site between LOS4-FITC and Bifidobacterium adolescentis was located outside and inside of cell, which was completed within 1 h. The relationship between structures of LOSs and prebiotics of intestinal flora (especially Bifidobacterium), and expanded the knowledge on the effects of carbohydrate polymerization degree (DP) and glycosidic bond connection with fermentation selectivity of bacteria was studied. [ABSTRACT FROM AUTHOR]

• Human milk N -glycome is utilized intracellularly by B. infantis ATCC 15697. • A novel strategy was setup for principle functional N -glycan identification. • Galactose-containing glycans were the principle bifidogenic components. Human milk N -glycome promotes the growth of Bifidobacterium longum subsp. infantis ATCC 15697. However, the action mode of, and the major functional components for, the bifidogenic function of human milk N -glycome remain unclear. In this study, we demonstrated that milk N -glycome was transferred in an intact form from culture into the bacterial cell and then decomposed intracellularly, evidenced by the following facts: (1) No UHPLC peak shift of N -glycome recovered from culture was observed. (2) No milk N -glycan specific monosugar was detected in culture supernatant. (3) High intracellular exoglycosidase activities were detected. (4) Fluorescently labeled N -glycans were found to be located intracellularly using Laser Scanning Confocal Microscopy (LSCM). Regarding the principal components identification, a novel sequential deglycosylation-based strategy was established. Degalactosylation, defucosylation-desialylation, and defucosylation-desialylation-degalactosylation treatments of human milk N -glycome showed that galactose-containing glycans were the principal components for the probiotic function of human milk N -glycome towards B. infantis ATCC 15697. [ABSTRACT FROM AUTHOR]

Human milk oligosaccharides (HMOs) are a family of structurally diverse unconjugated glycans that exhibit a wide range of biological activities. In this report, we describe an efficient, Multi-Enzyme One-Pot strategy to produce HMO mimics by the sialylation of galacto-oligosaccharides (GOSs), which are often added to infant formula as an inexpensive alternative to HMOs. In this system, the sialyltransferase donor, cytidine-5′-monophospho-N-acetylneuraminic acid (CMP-Neu5Ac), was generated in situ using a CMP-sialic acid synthetase. The sialylated GOSs were obtained by one-step purification after digesting CMP using the alkaline phosphatase PhoA to cytidine and inorganic phosphate. Although the synthesized α2,3-, α2,6- and α2,3/8-sialyl-GOSs exhibit different sialylation levels and patterns, all of these mixtures can be fermented by Bifidobacterium longum subsp. infantis ATCC 15697 but not by Bifidobacterium adolescentis ATCC 15703. The sialidase NanH2, which is unique to the former strain, hydrolyzed all of the synthesized HMO mimics. [ABSTRACT FROM AUTHOR]

Abstract: The effect of a yoghurt supplement containing Bifidobacterium pseudocatenulatum G4 or Bifidobacterium longum BB536 on plasma lipids, lipid peroxidation and the faecal excretion of bile acids was examined in rats fed a cholesterol-enriched diet. After 8weeks, the rats in the positive control (PC) group who were fed the cholesterol-enriched diet showed significant increases in plasma total cholesterol (TC), low-density lipoprotein (LDL) cholesterol, and malondialdehyde (MDA). However, groups fed a cholesterol-enriched diet supplemented with yoghurt containing B. pseudocatenulatum G4 or B. longum BB536 had significantly lower plasma TC, LDL-C, very-low-density lipoprotein (VLDL) cholesterol, and MDA than had the PC group after 8weeks of treatment. In addition, faecal excretion of bile acids was markedly increased in the rats fed the yoghurt containing B. pseudocatenulatum G4 or B. longum BB536 as compared to the PC and NC groups. [Copyright &y& Elsevier]

Abstract: Polyphenolic compounds, such as caffeic acid, offer many health benefits, but their industrial applications are limited because of their low solubility in water and instability under UV light or heat. In this study, we enzymatically produced caffeic acid glucosides, in both aqueous buffer and aqueous-supercritical carbon dioxide (SC-CO2) media, using a recombinant sucrose phosphorylase (SPase) from Bifidobacterium longum. By using LC/MS/MS analysis, we verified that the enzymatic reaction products were caffeic acid monoglucosides and diglucosides. Under SC-CO2, the amounts of the reaction products from the enzyme reaction were smaller than those in the aqueous reaction medium. However, this is the first report of the transglucosylation of caffeic acid by SPase, and also the first enzymatic reaction with phenolic compounds conducted in a SC-CO2 phase. [Copyright &y& Elsevier]

• Three LAMP primer sets were newly designed to distinguish B. longum subspecies. • A direct DNA extraction method was optimized to be suitable for probiotic products. • LAMP assays were combined with direct extraction method for on-site detection. • Direct LAMP assays successfully detected targets in probiotic products within 45 min. • This is the first study to identify B. longum subspecies using direct LAMP assay. As interest in probiotics increases, the need for accurate description of probiotic compositions present in commercial products is also increasing. Since Bifidobacterium longum used as probiotics is labeled at species or subspecies levels, a detection method for distinguishing B. longum subsp. longum, infantis , and suis is needed. Thus, we designed three LAMP primer sets for B. longum subspecies. Each primer set was specific for the target subspecies. The detection level was 0.2 pg for each target DNA (about 102 CFU/mL). To apply these LAMP assays to on-site detection, a direct DNA extraction method was optimized and combined with LAMP assay. Finally, direct LAMP assays were used to monitor the presence of B. longum subspecies in 16 probiotic products. They could specifically and sensitively detect target subspecies within approximately 45 min. These rapid on-site detection methods are useful for identifying B. longum subspecies in probiotic products. [ABSTRACT FROM AUTHOR]

• BlBE was overexpressed in Escherichia coli using the BLMA promoter. • BlBE was heterologous to known allergens and cold-active even at 10 °C. • BlBE preferred to transfer maltohexaose and introduced DP 4–36 branches into amylose. • BlBE increased the proportion of DP 2–10 branches in amylopectin and decreased its MW. • BlBE reduced the retrogradation and enhanced the slow digestibility of wheat starch. To develop a 1,4-α-glucan branching enzyme (BE) without homology to known allergens, the glgB gene from Bifidobacterium longum was overexpressed under the control of BLMA promoter in Escherichia coli. B. longum BE (BlBE) had a molecular weight of 86.1 kDa and a specific activity of more than 18.5 U/mg protein at 25–35 °C and pH 5.5–7.0, and exhibited 30% of the maximum activity at 10 °C. The cold-active BlBE preferred to transfer maltohexaose and introduced DP 4–36 branches into amylose. BlBE also increased the proportion of DP 2–10 branches in amylopectin and decreased its Mw from 1.39 × 106 to 1.16 × 105 g/mol. As the BlBE concentration increased from 0.0 to 0.5 U/mg substrate, the retrogradation enthalpy of BlBE-modified wheat starch decreased from 4.50 to 1.83 J/g (p < 0.05) at day 14 and the slowly digestible starch content increased from 2.10% to 17.39% (p < 0.05). [ABSTRACT FROM AUTHOR]