Verbascose

The Pea Oligosaccharides Could Stimulate the In Vitro Proliferation of Beneficial Bacteria and Enhance Anti-Inflammatory Effects via the NF-kappaB Pathway.[Pubmed:38397603]

Foods. 2024 Feb 19;13(4):626.

The oligosaccharides extracted from the seeds of peas, specifically consisting of raffinose, stachyose, and Verbascose, fall under the category of raffinose family oligosaccharides (RFOs). The effect of RFOs on intestinal microflora and the anti-inflammatory mechanism were investigated by in vitro fermentation and cell experiments. Firstly, mouse feces were fermented in vitro and different doses of RFOs (0~2%) were added to determine the changes in the representative bacterial community, PH, and short-chain fatty acids in the fermentation solution during the fermentation period. The probiotic index was used to evaluate the probiotic proliferation effect of RFOs and the optimal group was selected for 16S rRNA assay with blank group. Then, the effects of RFOs on the inflammatory response of macrophage RAW264.7 induced by LPS were studied. The activity of cells, the levels of NO, ROS, inflammatory factors, and the expression of NF-kappaB, p65, and iNOS proteins in related pathways were measured. The results demonstrated that RFOs exerted a stimulatory effect on the proliferation of beneficial bacteria while concurrently inhibiting the growth of harmful bacteria. Moreover, RFOs significantly enhanced the diversity of intestinal flora and reduced the ratio of Firmicutes-to-Bacteroides (F/B). Importantly, it was observed that RFOs effectively suppressed NO and ROS levels, as well as inflammatory cytokine release and expression of NF-kappaB, p65, and iNOS proteins. These findings highlight the potential of RFOs in promoting intestinal health and ameliorating intestinal inflammation.

The effect of seed germination and Bacillus spp. on the ripening of plant cheese analogs.[Pubmed:38319095]

Appl Environ Microbiol. 2024 Mar 20;90(3):e0227623.

Consumer demand for plant cheeses is increasing, but challenges of improving both flavor and quality remain. This study investigated the microbiological and physicochemical impact of seed germination and fermentation with Bacillus velezensis and Bacillus amyloliquefaciens on the ripening of plant cheese analogs. Chlorine treatment or addition of Lactiplantibacillus plantarum and Lactococcus lactis controlled microbial growth during seed germination. Lp. plantarum and Lc. lactis also served as starter cultures for the acidification of soy and lupine milk and were subsequently present in the unripened plant cheese as dominant microbes. Acidification also inhibited the growth and metabolic activity of bacilli but Bacillus spores remained viable throughout ripening. During plant cheese ripening, Lc. lactis was inactivated before Lp. plantarum and the presence of bacilli during seed germination delayed Lc. lactis inactivation. Metagenomic sequencing of full-length 16S rRNA gene amplicons confirmed that the relative abundance of the inoculated strains in each ripened cheese sample exceeded 99%. Oligosaccharides including raffinose, stachyose, and Verbascose were rapidly depleted in the initial stage of ripening. Both germination and the presence of bacilli during seed germination had impact on polysaccharide hydrolysis during ripening. Bacilli but not seed germination enhanced proteolysis of plant cheese during ripening. In conclusion, the use of germination with lactic acid bacteria in combination with Bacillus spp. exhibited the potential to improve the quality of ripened plant cheeses with a positive effect on the reduction of hygienic risks. IMPORTANCE: The development of novel plant-based fermented food products for which no traditional templates exist requires the development of starter cultures. Although the principles of microbial flavor formation in plant-based analogs partially overlap with dairy fermentations, the composition of the raw materials and thus likely the selective pressure on the activity of starter cultures differs. Experiments that are described in this study explored the use of seed germination, the use of lactic acid bacteria, and the use of bacilli to reduce hygienic risks, to acidify plant milk, and to generate taste-active compounds through proteolysis and fermentative conversion of carbohydrates. The characterization of fermentation microbiota by culture-dependent and culture-independent methods also confirmed that the starter cultures used were able to control microbial communities throughout 90 d of ripening. Taken together, the results provide novel tools for the development of plant-based analogs of fermented dairy products.

Isolation and structural elucidation of prebiotic oligosaccharides from Ziziphi Spinosae Semen.[Pubmed:37783055]

Carbohydr Res. 2023 Dec;534:108948.

Six oligosaccharides were discovered and isolated for the first time from Ziziphi Spinosae Semen. On the basis of spectroscopic analysis, their structures were determined to be Verbascose (1), verbascotetraose (2), stachyose (3), manninotriose (4), raffinose (5), and melibiose (6). The prebiotic effect of the oligosaccharide fraction was assayed by eight gut bacterial growth in vitro, revealing a significant increase in cell density, up to 4-fold, for Lactobacillus acidophilus, Lactobacillus gasseri, and Lactobacillus johnsonii. The impact of six oligosaccharides with different degrees of polymerization (DPs) and structures on the growth of Lactobacillus acidophilus was evaluated. As a result, stachyose and raffinose demonstrated superior support for bacterial growth compared to the other oligosaccharides. This study explored the structure-activity relationship of raffinose family oligosaccharides (RFOs) and showed that the more the monosaccharide type, the more supportive the gut bacteria growth when oligosaccharides have the same molecular weight.

The effects of microbial fertilizers application on growth, yield and some biochemical changes in the leaves and seeds of guar (Cyamopsis tetragonoloba L.).[Pubmed:37689887]

Food Res Int. 2023 Oct;172:113122.

Guar (Cyamopsis tetragonoloba L.) is a summer legume that is becoming a crucial industrial crop because of its high gum and protein content. Thus far, the combined effects of arbuscular mycorrhizal fungi (AMF) and Bradyrhizobium on the yield and chemical composition of guar plants are not well studied. Therefore, the current investigation was designed to estimate the individual as well as the combined effects of AMF and Bradyrhizobium on plant growth, yield and nutritional quality of seeds and leaves of guar. AMF and/or Bradyrhizobium inoculation improved chemical composition of guar seeds and its morpho-physiological (plant height, fresh weight, dry weight, and yield production) traits. In addition to increased guar growth and yield production, the inoculation of AMF and/or Bradyrhizobium increased guar leaf and seed minerals, fiber, lipids, crude protein and ash contents. At primary metabolites, there were increases in sugar levels including raffinose stachyose, Verbascose and galactomannan. These increases in sugar provided a route for organic acids, amino acids and fatty acids production. Interestingly, there was an increase in essential amino acids and unsaturated fatty acids. At the bioactive secondary metabolite levels, biofertilizers improved phenols and flavonoids levels and anthocyanin and polyamines biosynthesis. In line with these increases, precursors of anthocyanin (phenylalanine, p-coumaric acid, and cinnamic acid) and the levels of polyamines (diaminopropane, putrescine, cadaverine, spermidine, spermine, and agmatine) were increased. Overall, for the first time, our study shed the light on how AMF and Bradyrhizobium improved guar yield and metabolism. Our findings suggested that the combined inoculation of AMF and Bradyrhizobium is an innovative approach to improve guar growth, yield production and yield quality.

Characterization of free oligosaccharides from garden cress seed aqueous exudate using PGC LC-MS/MS and NMR spectroscopy.[Pubmed:37541111]

Carbohydr Res. 2023 Oct;532:108914.

Garden cress seeds produces mucilage that has found various food applications, however, there is little information on the free oligosaccharides (FOS) contents in these seeds. Herein, we explored the presence of FOS in cress seed aqueous exudate. PGC-LC MS/MS analysis indicated the presence of mainly hexose containing oligosaccharides such as raffinose, stachyose and Verbascose belonging to raffinose family of oligosaccharides (RFOs). In addition, minor fraction of planteose, isomeric tri- and tetrasaccharides were also observed. Further, the structural confirmation of the abundant tri- and tetrasaccharide were obtained through 1D and 2D NMR analysis. Thus, the RFOs presence in cress seeds would enhance its bio-functionalities.

Seven new pentasaccharides from the roots of Rehmannia glutinosa.[Pubmed:36877100]

J Asian Nat Prod Res. 2024 Feb;26(2):280-292.

Seven new pentasaccharides (1-7), rehmaglupentasaccharides A-G, were isolated from the air-dried roots of Rehmannia glutinosa. Their structures were established from the spectroscopic data obtained and by chemical evidence. The known Verbascose (8) and stachyose (9) were also obtained in the current investigation, and the structure of stachyose was unequivocally defined using X-ray diffraction data. Compounds 1-9 were tested for their cytotoxicity against five human tumor cell lines, influence on dopamine receptor activation, and proliferation effects against Lactobacillus reuteri.

The potential role of nondigestible Raffinose family oligosaccharides as prebiotics.[Pubmed:36795047]

Glycobiology. 2023 May 17;33(4):274-288.

Based on factual scientific health claims, prebiotics have gained significant importance in ever-growing food and pharmaceutical industries. The diverse nature of distinct prebiotics influences the host differently in distinguishable patterns. Functional oligosaccharides are either plant-derived or commercially prepared. Raffinose, stachyose, and Verbascose are the 3 types of raffinose family oligosaccharides (RFOs) that have been extensively used as medicine, cosmetic, and food additives. These dietary fiber fractions avert the adhesion and colonization by enteric pathogens and add nutrition metabolites for a healthy immune system. Enrichment of RFOs in healthy foods should be promoted as these oligosaccharides augment gut microecology by enhancing the health conferring microbes i.e. bifidobacteria and lactobacilli. RFOs influence the host's multiorgan systems due to their physiological and physicochemical properties. For example, the fermented microbial products of such carbohydrates affect neurological processes, including memory, mood, and behavior in humans. Raffinose-type sugar uptake is thought to be a ubiquitous property of bifidobacteria. This review paper summarizes the source of RFOs and their metabolizing entities, highlighting bifidobacterial carbohydrate utilization and health benefits.

Comprehensive oligosaccharide profiling of commercial almond milk, soy milk, and soy flour.[Pubmed:36586264]

Food Chem. 2023 May 30;409:135267.

Oligosaccharides are known for several bioactivities on health, however, in sensitive individuals, can cause intestinal discomfort. This study aimed to investigate the oligosaccharide profiles in selected plant-based food products. A quantification method based on high-performance anion-exchange chromatography-pulsed amperometric detection was developed, validated, and used to measure major oligosaccharides. Additional low-abundant oligosaccharides and glycosides were characterized by liquid chromatography-tandem mass spectrometry and glycosidases. The summed concentration of raffinose, stachyose, and Verbascose ranged from 0.12-0.19 mg/g in almond milk, 3.6-6.4 mg/g in soy milk, and 74-77 and 4.8-57 mg/g in defatted and full-fat soy four. Over 80 different oligosaccharides were characterized. Novel compounds, 2,3-butanediol glycosides, were identified in almond milk. Low-abundant oligosaccharides represented 25 %, 6 %, and 10 % of total OS in almond milk, soy milk, and soy flour, respectively. The data here are useful to estimate oligosaccharide consumption from dietary intake and facilitate further studies on their bioactivity.

Assessment of diversity in anti-nutrient profile, resistant starch, minerals and carbohydrate components in different ricebean (Vigna umbellata) accessions.[Pubmed:36356361]

Food Chem. 2023 Mar 30;405(Pt A):134835.

Ricebean accessions (n = 38) cultivated in India were evaluated for their comprehensive nutrient, anti-nutrients and mineral composition. Protein and total dietary fibre ranged between 23.23 and 27.33 and 12.27 to 16.69 g/100 g, respectively. Among the oligosaccharides, Verbascose was not detected, however, raffinose and stachyose ranged between 47 and 186 and 117 to 5765 mg/100 g, respectively. Among the free sugars, sucrose was found dominating (up to 370 mg/100 g). Resistant starch (4.13 to 8.62 %), iron (3.49 to 7.46 mg/100 g), zinc (1.90 to 3.72 mg/100 g) and selenium (0.28 to 4.48 microg/100 g) varied significantly (p < 0.05) among ricebean samples. Phytic acid, saponin, trypsin inhibitor and oxalate analysed in ricebean accessions ranged between 303 and 760 mg/100 g, 19 to 46 mg/g, 309 to 1076 mg/100 g and 219 to 431 mg/100 g, respectively. Multivariate analysis using hierarchical clustering analysis (HCA), and principal component analysis (PCA) was employed to decipher the diversity of nutrients and anti-nutrients across the ricebean accessions. Based on HCA, dendrogram-1 (nutrients) and dendrogram-2 (minerals, anti-nutrients) were produced, having four clusters in each. In the dendrogram-1 and 2, the largest cluster had (n = 21) and (n = 15) accessions, respectively. The PCA analyse the uncorrelated set of variables (principal components) and it condenses a large set of data variables. Based on the eigenvalue >1, a total of eight PCs were formed contributing total variance of 78.8 %. The factor loading contribution in the PC1 and PC2 were from iron, fructose, glucose, raffinose and total dietary fibre, selenium (Se) and protein, respectively.

Isolation and Identification of an alpha-Galactosidase-Producing Lactosphaera pasteurii Strain and Its Enzymatic Expression Analysis.[Pubmed:36144675]

Molecules. 2022 Sep 13;27(18):5942.

alpha-Galactosidase (EC 3.2.1.22) refers to a group of enzymes that hydrolyze oligosaccharides containing alpha-galactoside-banded glycosides, such as stachyose, raffinose, and Verbascose. These enzymes also possess great potential for application in sugar production, and in the feed and pharmaceutical industries. In this study, a strain of Lactosphaera pasteurii (WHPC005) that produces alpha-galactosidase was identified from the soil of Western Hunan, China. It was determined that the optimal temperature and pH for this alpha-galactosidase were 45 degrees C and 5.5, respectively. The activity of alpha-galactosidase was inhibited by K(+), Al(3+), Fe(3+), fructose, sucrose, lactose, galactose, SDS, EDTA, NaCl, and (NH(4))(2)SO(4), and enhanced by Ca(2+), Fe(2+), Mn(2), Zn(2+), glucose, and raffinose. The optimal inducer was raffinose, and the optimal induction concentration was 30 mumol/L. The alpha-galactosidase gene was cloned using random fragment cloning methods. Sequence analysis demonstrated that the open reading frame of the alpha-galactosidase gene was 1230 bp, which encodes a putative protein of 409 amino acids in length. Bioinformatics analysis showed that the isoelectric point and molecular weight of this alpha-galactosidase were 4.84 and 47.40 kD, respectively. Random coils, alpha helixes, and beta turns were observed in its secondary structure, and conserved regions were found in the tertiary structure of this alpha-galactosidase. Therefore, this alpha-galactosidase-producing bacterial strain has the potential for application in the feed industry.

Structural Characterization of Dietary Fiber from Different Lupin Species (Lupinus sp.).[Pubmed:35758602]

J Agric Food Chem. 2022 Jul 13;70(27):8430-8440.

Dietary fiber fractions of whole seeds from different lupin species were structurally characterized. The low-molecular-weight soluble dietary fiber fraction contains mainly stachyose and Verbascose. The soluble dietary fiber fraction is dominated by homogalacturonan and rhamnogalacturonan type I (RGI), with (arabino-)galactans and to a lesser portion arabinans as neutral RGI side chains. Arabinans are preferentially branched in position O2 as demonstrated by methylation analysis and an arabinan profiling approach. Insoluble dietary fiber is mainly composed of cellulose and pectins, but xylans and xyloglucans are present, too. Application of an enzymatic xyloglucan profiling approach demonstrated a substitution degree of 75% and proved the existence of fucosylated xyloglucans. Lignin of all lupin species was analyzed as being rich in guaiacyl units; however, the degree of lignification is low. Alcohol-insoluble residue polysaccharides from both seed coat and embryo/endosperm were analyzed separately, demonstrating tissue-related differences in the portions of cellulose and RGI.

An allelic series of starch-branching enzyme mutants in pea (Pisum sativum L.) reveals complex relationships with seed starch phenotypes.[Pubmed:35450647]

Carbohydr Polym. 2022 Jul 15;288:119386.

A set of mutant pea lines carrying induced mutations within the major seed-expressed starch-branching enzyme gene has been characterised at the molecular, chemical and agronomic levels. Eight of the induced mutations, three of which predicted a premature stop codon, were compared with the naturally occurring starch-branching enzyme mutation within the same genetic background. Starch, amylose and sugar measurements, coupled with analysis by ultra-high performance liquid chromatography-size exclusion chromatography of starches, identified a range of phenotypes which were grouped according to the nature of the mutation. Homology modelling of proteins supported the differences in phenotypes observed. Differences in field performance were evident for selected mutants, particularly in seed yield and mean seed weight traits for early compared with late spring sowings. The data show the potential of an allelic series of mutants at this locus for nutritional studies. CHEMICAL COMPOUNDS: starch, amylose, amylopectin, raffinose, stachyose, Verbascose.

The Effectiveness of Extruded-Cooked Lentil Flour in Preparing a Gluten-Free Pizza with Improved Nutritional Features and a Good Sensory Quality.[Pubmed:35159632]

Foods. 2022 Feb 7;11(3):482.

Extruded-cooked lentil (ECL) flour was used to fortify (10/100 g dough) gluten-free pizza, which was compared with rice/corn-based pizza (control), and with pizza containing native lentil (NL) flour. Viscoamylograph and Mixolab data evidenced the hydrocolloid properties of ECL flour (initial viscosity = 69.3 BU), which contained pregelatinized starch. The use of ECL flour made it possible to eliminate hydroxymethylcellulose (E464), obtaining a clean label product. Both NL and ECL pizzas showed significantly (p < 0.05) higher contents of proteins (7.4 and 7.3/100 g, respectively) than the control pizza (4.4/100 g) and could be labelled as "source of proteins" according to the Regulation (EC) No. 1924/2006. In addition, NL and ECL pizzas were characterized by higher contents of bioactive compounds, including anthocyanins, and by higher in vitro antioxidant activity (1.42 and 1.35 micromol Trolox/g d.m., respectively) than the control pizza (1.07 micromol Trolox/g d.m.). However, NL and ECL pizzas also contained small amounts of undigestible oligosaccharides, typically present in lentils (Verbascose = 0.92-0.98 mg/g d.m.; stachyose = 4.04-5.55 mg/g d.m.; and raffinose = 1.98-2.05 mg/g d.m.). No significant differences were observed in the liking level expressed by consumers between ECL and control pizzas.

Sequential expression of raffinose synthase and stachyose synthase corresponds to successive accumulation of raffinose, stachyose and verbascose in developing seeds of Lens culinaris Medik.[Pubmed:34454370]

J Plant Physiol. 2021 Oct;265:153494.

Raffinose, stachyose and Verbascose form the three major members of the raffinose family oligosaccharides (RFO) accumulated during seed development. Raffinose synthase (RS; EC 2.4.1.82) and stachyose synthase (STS; EC 2.4.1.67) have been associated with raffinose and stachyose synthesis, but the precise mechanism for Verbascose synthesis is not well understood. In this study, full-length RS (2.7 kb) and STS (2.6 kb) clones were isolated by screening a cDNA library prepared from developing lentil seeds (18, 20, 22 and 24 days after flowering [DAF]) to understand the roles of RS and STS in RFO accumulation in developing lentil seeds. The nucleotide sequences of RS and STS genes were similar to those reported for Pisum sativum. Patterns of transcript accumulation, enzyme activities and RFO concentrations were also comparable to P. sativum. However, during lentil seed development raffinose, stachyose and Verbascose accumulation corresponded to transcript accumulation for RS and STS, with peak transcript abundance occurring at about 22-24 DAF, generally followed by a sequential increase in raffinose, stachyose and Verbascose concentrations followed by a steady level thereafter. Enzyme activities for RS, STS and Verbascose synthase (VS) also indicated a sudden increase at around 24-26 DAF, but with an abrupt decline again coinciding with the subsequent steady state increase in the RFO. Galactan:galactan galactosyl transferase (GGT), the galactinol-independent pathway enzyme, however, exhibited steady increase in activity from 24 DAF onwards before abruptly decreasing at 34 DAF. Although GGT activity was detected, isolation of a GGT sequence from the cDNA library was not successful.

Multiple reaction monitoring for identification and quantification of oligosaccharides in legumes using a triple quadrupole mass spectrometer.[Pubmed:34392119]

Food Chem. 2022 Jan 30;368:130761.

Raffinose family oligosaccharides are non-digestible compounds considered as dietary prebiotics with health-related properties. Hence, it is important to develop highly specific methods for their determination. An analytical method is developed in this study for oligosaccharide identification and quantification using liquid chromatography-tandem mass spectrometry equipped with a triple quadrupole analyser operating in Multiple Reaction Monitoring mode. Raffinose, stachyose and Verbascose are separated in a 10-minute run and the method is validated over a broad concentration range, showing good linearity, accuracy, precision and high sensitivity. A low-cost, short eco-friendly procedure for oligosaccharide extraction from legumes, with a high recovery rate extraction, good repeatability and reproducibility is also proposed. No plant-matrix effects were demonstrated. The method applied to the screening of 28 different legumes revealed species-related traits for oligosaccharide distribution, highlighting Pisum sativum (9.22 g/100 g) as the richest source of these prebiotics and its suitability as a functional food ingredient.