Search Results (63)

Fructooligosaccharides (FOS) are short fructans with different degrees of polymerization (DP) and bonds in their structure, generated by the distinct activities of fructosyltransferase enzymes, which produce distinct types of links. FOS are in high demand on the market, mainly because of their prebiotic effects. In recent years, depending on the link type in the FOS structure, prebiotic activity has been shown to be increased. Studies on β-fructanofuranosidases (Ffasa), enzymes with fructosyltransferase activity in yeasts, have reported the production of ¹F-FOS, ⁶F-FOS, and ⁶G-FOS. The aims of this investigation were to evaluate the capability of fifteen different yeasts to grow in Agave sp. juices and to determine the potential of these juices as substrates for FOS production. Additionally, the research aimed to corroborate and analyze the fructosyltransferase activity of enzymatic extracts obtained from agave yeasts by distinct induction media and to identify the role and optimal parameters (time and sucrose and glucose concentrations) for FOS and disaccharides production through Box–Behnken designs. To carry out such a task, different techniques were employed: FT-IR, TLC, and HPAEC-PAD. We found two yeasts with fructosyltransferase activity, P. kudriavzevii ITMLB97 and C. lusitaniae ITMLB85. In addition, within the most relevant results, the production of the FOS 1-kestose, 6-kestose, and neokestose, as well as disaccharides inulobiose, levanobiose, and blastose, molecules with potential applications, was determined. Overall, FOS production requires suitable yeast species, which grow in a medium under optimal conditions, from which microbial enzymes with industrial potential can be obtained. Full article

Background: Prebiotics are non-digestible dietary compounds, defined as substrates that are utilised by host microorganisms conferring a health benefit. Although fructo-oligosaccharides (FOSs) and galacto-oligosaccharides (GOSs) are among the most studied prebiotics and support intestinal normobiosis, comprehensive data on their content in foods remain limited. Objectives: The objective was to quantify the content of FOSs (kestose, nystose, and 1 F-β-fructofuranosylnystose) and GOSs (raffinose and stachyose) in 35 foods, including fruit and nuts, legumes, and cereals. We also estimated the intakes of prebiotics in an Italian population. Methods: We analysed the prebiotic content in foods using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). We estimated the prebiotic intake of 100 healthy controls from a case-control study on colorectal cancer conducted in Italy between 2017 and 2019. We used dietary information collected through a food frequency questionnaire and the prebiotic data quantified in this and a previous study. Results: FOSs were mostly detected in cereal products, with wheat bran and whole-meal rye flour containing the highest amount (around 0.7 g/100 g each). GOSs were most abundant in legumes, especially in dried soy products (around 4.0 g/100 g each). Mean daily intake was 0.236 g for total FOSs and 0.371 g for total GOSs. Wheat bran, raspberries, chestnuts, walnuts, raisins, soy milk, and soy yoghurt overall accounted for 3.9% of kestose, 1.2% of nystose, 0% of 1F-β-fructofuranosylnystose, 15.5% of raffinose, and 8.3% of stachyose total intakes. Conclusions: The present study enables the development of a comprehensive database on prebiotic content in foods through a consistent analytical method. This makes prebiotic intake assessments more accurate than previously available data and facilitates future epidemiological studies investigating their potential effects on health. Full article

Objectives: The purpose of this study was to further characterize the ultrafiltration (UF) method for determining free saccharide levels in glycoconjugate vaccines and compare it with other methods used for the determination of free saccharide levels in meningococcal glycoconjugate vaccines. Methods: We performed experiments on both meningococcal glycoconjugates and capsular polysaccharides, and compared UF, deoxycholate (DOC) precipitation, and solid-phase extraction (SPE) methods. Meningococcal capsular polysaccharides from groups A (MenA), C (MenC), and W (MenW) were depolymerized and characterized using SEC-MALS (size-exclusion chromatography with multi-angle laser light scattering) to determine the molecular weight and hydrodynamic size and then subjected to UF. The free saccharide content was quantified using HPAEC-PAD (high-performance anion-exchange chromatography with pulsed amperometric detection). Results: The characterization of size-reduced group C polysaccharide revealed weight-average molecular mass (Mw) ranging from 22,200 g/mol to 287,300 g/mol and hydrodynamic radii of 3.7 to 19.5 nm. Pore size studies confirmed that polysaccharides with diameters up to 15 nm filtered through the 100 kDa cellulose membrane. The smallest PS fragment tested (22,200 g/mol, 7.4 nm diameter) was partially recovered from the 30 kDa membrane. For MenC-CRM₁₉₇, DOC yielded the lowest free saccharide content (<1%), UF gave moderate results (7–8%), and SPE showed the highest and most variable values (up to 15%). For MenA- and MenW-CRM₁₉₇, UF and DOC consistently provided low free saccharide levels (<2% and 3–11%, respectively). Conclusions: The upper limits on the size of free group C meningococcal polysaccharides that can be ultrafiltered were assessed. Differences in the relative amount of free saccharide were observed between various methods used to control meningococcal conjugate vaccines. Full article

The Cucurbita genus comprises various species that are globally consumed and that are commonly used for their nutritional value but also for medicinal applications. Within the Cucurbita genus can be found Cucurbita ficifolia Bouché, a species that is understudied regarding its potential value for the food industry, as a functional food, and for the pharmaceutical industry, as a source of nutraceuticals. Therefore, in this study we investigated the phytochemical composition and bioactivities of aqueous (AF) and methanolic (MF) fractions of C. ficifolia pulp and peel hydroethanolic (HE) extracts. HPLC-DAD-MSⁿ and HPAEC-PAD analyses of extracts’ fractions revealed a low content of polyphenols and a significant content of sugars. Through in vitro inhibition assays of the enzymes alpha-amylase, acetylcholinesterase (AChE), and elastase, all fractions showed, respectively, antidiabetic, neuroprotective, and anti-aging activities. The safety profile and anti-tumoral activities were evaluated in various cell models (Caco-2, HaCaT, HepG2, and RAW 264.7), and results showed that the fractions obtained from pulp extract induce no/low cytotoxicity, while the methanolic fraction of peel induced cytotoxicity in all cell lines. At non-cytotoxic concentrations, aqueous and methanolic fractions of both extracts significantly inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, revealing anti-inflammatory activity. Flow cytometry analysis showed that both aqueous fractions increased basal levels of glutathione (GSH) in Caco-2 cells, while not inducing oxidative stress, revealing potential as antioxidant dietary agents. However, the MF of peel HE extract induced oxidative stress in Caco-2 cells, as it increased reactive oxygen species (ROS) and lipid peroxidation. AF fraction of peel extract induced cell cycle arrest in the G0/G1 phase, while the other fractions induced cell cycle arrest in the S phase. In conclusion, Cucurbita ficifolia fruit presents potential as a functional food but also as a potential source of nutraceuticals, and peel waste products can be valorized by pharmaceutical and cosmeceutical industries as sources of bioactive molecules. Full article

This study assessed the technological potential and bioactive compounds present in purple yampee (Dioscorea trifida L.f.) lyophilized powder, peeled and whole flour, as well as the tuber peel, starch residual fiber, and wastewater mucilage. Although most values approached neutrality, flour showed a lower pH and high density, while greater acidity was observed in the mucilage. Color differed statistically and perceptibly between all samples, with similar values of °hue to purple flours from other sources, and the maximum chroma was found in lyophilized pulp and lightness in fiber. Average moisture levels around 7.2% and water activity levels of 0.303 (0.194 for whole flour) in fractions suggested favorable storability, while the interaction of the powders with water was similar to other root and tuber powdered derivatives. Yampee periderm had the highest swelling power, oil absorption capacity, water holding capacity, and absorption index and capacity. Mucilage had a higher solubility index and outstanding emulsion activity, greater than 90%. Twelve anthocyanins, with new reports of petunidin derivatives for the species, and more than 30 phytochemicals were identified through advanced liquid chromatography techniques. The greatest amounts of pinitol and myo-inositol were found in the mucilage, and sucrose, glucose, and fructose prevailed in the other powders. Successfully characterized yampee fractions showed high potential as functional food ingredients. Full article

Single enzymatic modifications are limited to starch. Complex modification with synergistic amylases will improve starch properties more significantly. In this study, maize starch was compound modified by β-amylase and α-glucosidase. The structure and physicochemical properties of the corn starch were determined by scanning electron microscopy (SEM), X-ray diffractometry (XRD), Fourier transform infrared spectroscopy (FT–IR), proton nuclear magnetic resonance hydrogen spectroscopy (¹HNMR), high-performance anion-exchange chromatography (HPAEC–PAD), differential scanning calorimetry (DSC) and Rapid Visco analyzer (RVA) to determine the changes in the structure and physicochemical properties of maize starch before and after the dual enzyme modification. The branching degree (4.95–7.10%) of maize starch was increased after bi-enzymatic modification, the amylose content (28.77–18.60%) was decreased, and the amylopectin content (70.79–81.71%) was elevated. The relative crystallinity (20.41–30.20%) and short-range ordered structure of the starch increased, and the dual enzyme modification led to a more compact structure. Dual enzyme-modified maize starch showed a decrease in long chains, an increase in short chains, and its degree of branching was elevated. Dual enzyme modification also affected the thermal stability, pasting, light transmittance (1.40–2.16%), solubility (20.15–13.76%), and swelling (33.97–45.79%) of maize starch. It can be concluded that the complex modification of maize starch by β-amylase and α-glucosidase significantly changed the amylose/amylopectin ratio of the starch and made its structure denser. These results can provide a theoretical basis for the enzymatic preparation of maize starch with different amylose/amylopectin ratios and the development and utilization of functional starches. Full article

Immobilized enzyme reactors (IMERs) are critical tools for developing novel oligosaccharides based on the enzymatic catalysis of polysaccharides. In this paper, a novel glucuronan lyase from Peteryoungia rosettiformans was produced, purified, and then immobilized on a CIM^® CDI disk for cleaving glucuronan. The results showed that around 63.6% of glycuronan lyases (800.9 μg) were immobilized on the disk. The V_max values of immobilized glucuronan lyases did not significantly change (56.9 ± 4.7 μM∙min⁻¹), while the K_m values (0.310 ± 0.075 g∙L⁻¹) increased by 2.5 times. It is worth noting that immobilized glucuronan lyases overcame the catalytic inhibition of free enzymes observed under high glucuronan concentrations (0.5–2 g∙L⁻¹). circumscribed central composite design (CCCD) and response surface methodology (RSM) showed that glucuronan concentration, flow rate, and reaction time significantly affected the yield of oligoglucuronans. The degree of polymerization (DP) of degraded glucuronan ranged from DP 2–8 according to the results obtained by high performance anion exchange chromatography coupled with a pulsed amperometric detector (HPAEC-PAD). The IMER retained 50.9% activity after running 2373 column volumes of glucuronan. Finally, this glucuronan lyase reactor was tentatively connected to an immobilized laccase reactor to depolymerize, and gallic acid (GA) was added to glucuronan. Approximately 8.5 mg of GA was added onto 1 g of initial glucuronan, and the GA–oligoglucuronan conjugates showed notable antioxidant activity. Full article

Obligate anaerobic beer spoilage bacteria have been a menace to the brewing industry for several decades. Technological advances in the brewing process aimed at suppressing aerobic spoilers gave rise to problems with obligate anaerobes. In previous studies, the metabolic spectrum of Pectinatus and Megasphaera species has been described, but their metabolism in the beer environment remains largely unknown. We used high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME-GCMS) to further characterize beer spoiled by 30 different strains from six beer-spoiling species of Pectinatus and Megasphaera (P. cerevisiiphilus, P. frisingensis, P. haikarae, M. cerevisiae, M. paucivorans, and M. sueciensis). We detected differences in carbohydrate utilization and the volatile organic compounds (volatilome) produced during beer spoilage by all six species. We were able to show that glycerol, one of the basic components of beer, is the common carbon source used by all strains. It appears that this carbon source allows for anaerobic beer spoilage by Pectinatus and Megasphaera despite the spoilage-preventing intrinsic barriers of beer (iso-α-acids, ethanol, low pH, scarce nutrients); thus, extrinsic countermeasures are key for prevention. Full article

Several chemical analytical methods were applied to characterize the chemical structure of polysaccharides extracted from discarded apples and pomegranate peels using hydrodynamic cavitation methods in a circular economy perspective. In particular, the purity of the polysaccharides and the degrees of acetylation and methylation were evaluated by proton Nuclear Magnetic Resonance (¹H-NMR) analysis; simple sugars and galacturonic acid were analyzed simultaneously by High-Performance Anion Exchange Chromatography—Pulsed Amperometric Detector (HPAEC-PAD); the molecular weight of the extracted polysaccharides was determined by High-Performance Size Exclusion Chromatography-Refractive Index Detector (HPSEC-RID). The results showed a negligible presence of co-precipitated proteins/tannins, easily removed by dialysis, as well as other co-precipitated molecules such as monosaccharides and organic acids. Polysaccharides from apples consisted mainly of pectic material with a prevalence of homogalacturonans. Polysaccharides from pomegranate peels showed greater compositional variability with significant amounts of arabinose and galactose, a lower content of pectin, and the presence of rhamnogalacturonans I. Both polysaccharides were highly methylated and differed in the degree of acetylation, which could lead to different properties. Polysaccharides from apples presented two main molecular weights (>805 kDa and 348–805 kDa, respectively), while those from pomegranate peel showed a major fraction at 348 kDa and minor fractions < 23 kDa. In conclusion, the research tools proposed by this study have allowed defining the macrostructure of polysaccharides in a quick and efficient way to valorize these food by-products. Full article

Acid hydrolysis serves as the primary method for determining the monosaccharide composition of polysaccharides. However, inappropriate acid hydrolysis conditions may catalyze the breakdown of monosaccharides such as fructans (Fru), generating non-sugar by-products that affect the accuracy of monosaccharide composition analysis. In this study, we determined the monosaccharide recovery rate and non-sugar by-product formation of inulin-type fructan (ITF) and Fru under varied acid hydrolysis conditions using HPAEC-PAD and UPLC-Triple-TOF/MS, respectively. The results revealed significant variations in the recovery rate of Fru within ITF under different hydrolysis conditions, while glucose remained relatively stable. Optimal hydrolysis conditions for achieving a relatively high monosaccharide recovery rate for ITF entailed 80 °C, 2 h, and 1 M sulfuric acid. Furthermore, we validated the stability of Fru during acid hydrolysis. The results indicated that Fru experienced significant degradation with an increasing temperature and acid concentration, with a pronounced decrease observed when the temperature exceeds 100 °C or the H₂SO₄ concentration surpasses 2 M. Finally, three common by-products associated with Fru degradation, namely 5-hydroxymethyl-2-furaldehyde, 5-methyl-2-furaldehyde, and furfural, were identified in both Fru and ITF hydrolysis processes. These findings revealed that the degradation of Fru under acidic conditions was a vital factor leading to inaccuracies in determining the Fru content during ITF monosaccharide analysis. Full article

Currently, the demand in the food market for oligosaccharides with biological activities is rapidly increasing. In this study, agar polysaccharides from Gracilaria fisheri were treated with β-agarases and hydrolyzed to agar oligosaccharides (AOSs). High-performance anion-exchange chromatography/pulsed amperometric detection (HPAEC-PAD), Fourier-transform infrared spectroscopy (FT-IR), and gel permeation chromatography (GPC), were employed to analyze the chemical characteristics of AOSs. The FT-IR spectra revealed that the enzymatic hydrolysis had no effect on specific functional groups in the AOS molecule. To investigate the prebiotic and pathogen inhibitory effects of AOSs, the influence of AOSs on the growth of three probiotic and two pathogenic bacteria was examined. The gastrointestinal tolerance of probiotics in the presence of AOSs was also investigated. AOSs enhanced the growth of Lactobacillus plantarum by 254%, and inhibited the growth of Bacillus cereus by 32.80%, and Escherichia coli by 58.94%. The highest survival rates of L. plantarum and L. acidophilus were maintained by AOSs in the presence of α-amylase and HCl under simulated gastrointestinal conditions. This study demonstrates that AOSs from G. fisheri exhibit potential as a prebiotic additive in foods. Full article

Fructans serve as the primary form of storage carbohydrate in cool-season grasses, but little is known about potential differences in ruminal fermentation of fructans between cattle and sheep. An ex vivo study was conducted to evaluate species differences in fructan catabolism. Buffered media containing ground orchardgrass (Dactylis glomerata L.) substrate was inoculated with uncultivated rumen microbiota obtained from cattle and sheep (n = 4 species⁻¹). Fructan profiles were monitored over the incubation period (8 h; 39 °C) using high-performance anion-exchange chromatography coupled to pulsed amperometric detection (HPAEC-PAD). In both species, disappearance of long-chain fructans (degree of polymerization [DP] > 8) was evident by 2 h of incubation (p < 0.01), whereas short-chain fructans (DP 4–8) increased from 0 to 2 h prior to subsequent degradation (p < 0.01). However, the overall rate of long-chain fructan catabolism was greater in bovine versus ovine fermentations, particularly between 2 and 4 h (p < 0.01). Additionally, rapid utilization of short-chain fructans occurred from 2 to 4 h in bovine fermentations, but was delayed in ovine fermentations, with substantial degradation occurring only after 4 h of incubation (p < 0.01). These results indicate that rumen microbiota of cattle may have a greater capacity for fructan degradation. Full article

The rhizome of Polygonatum, which has been consumed in China for nearly 2000 years, is rich in oligosaccharides, polysaccharides, saponins, and alkaloids and has various effects such as lowering blood sugar and anti-aging. Its aerial parts are often discarded as by-products and have been rarely studied; moreover, the nutritional composition and carbohydrate composition of these parts are not clear. In this study, the nutritional composition, amino acid evaluation, and the total phenolics content of the leaves, stems, fruits, and flowers of Polygonatum cyrtonema Hua were analyzed, and the composition and content of carbohydrates were determined by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC–PAD). The results suggested that the leaves and flowers of Polygonatum cyrtonema Hua (PC) contained 1.12–1.37% phenolic substances. The proteins of the aerial parts had high nutritional values, and the total amino acid content of the leaves and flowers, with the total leaf amino acids amounting to 11.77%, were followed by the flowers at 11.65%. The sugars in the flowers were mainly free monosaccharides at 10.38%, and the fruits were dominated by 9.44% fructo-oligosaccharides. This study provided new evidence for the utilization of the aerial parts of Polygonatum. Full article

A new skincare application scenario for dark tea, a unique and post-fermented tea popular in the health food industry, was developed in this paper. The effects of dark tea polysaccharide (DTP) on stress-induced skin problems and its mechanism of action were investigated by modeling cortisone-induced stress injury in human HaCaT keratinocytes and SZ95 sebaceous gland cells. The results showed a reduced cortisol conversion induced by cortisone under the action of DTP with a concentration of 200 μg/mL, probably by inhibiting the expression of the HSD11B1 enzyme. DTP was also able to suppress the cortisone-induced elevation of lipid levels in SZ95 sebocytes at this concentration. In addition, the composition and structure of DTP were verified by ultrafiltration, ultraviolet-visible spectrophotometry (UV-VIS), high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and infrared spectroscopy. In brief, DTP has a unique and significant stress-relieving effect, which provides new ideas for the development of new ingredients for the skin care industry. Full article

Background: “FODMAPs” (fermentable-oligo-, di-, monosaccharides, and polyols) are a group of fermentable carbohydrates and polyols largely diffused in food products. Despite their beneficial effects as prebiotics, people affected by irritable bowel syndrome manifest symptoms when eating these carbohydrates. A low-FODMAP diet seems to be the only possible therapy proposed for symptom management. Bakery products are a common source of FODMAPs, whose pattern and total amount can be affected by their processing. This work aims at studying some of the technological parameters that can influence the FODMAPs pattern in bakery products during the production process. Methods: high-performance anion exchange chromatography coupled to a pulsed amperometric detector (HPAEC-PAD) was used as a highly selective system for carbohydrates evaluation analyses on flours, doughs, and crackers. These analyses were performed using two different columns, the CarboPac PA200 and CarboPac PA1, which are selective for oligosaccharide and simple sugar separation, respectively. Results: emmer and hemp flours were selected to prepare doughs as they contained low oligosaccharide content. Two different mixes of ferments were used at different times of fermentation to evaluate the best conditions to achieve low-FODMAP crackers. Conclusion: the proposed approach allows carbohydrate evaluation during crackers processing and permits the selection of opportune conditions to obtain low-FODMAP products. Full article