RESUMO
Recently, there has been a burgeoning interest in harnessing the potential of biomass and industry byproducts for the development of novel products and materials. In particular, this study explored the efficient valorization of sunflower meal (SFM), an underutilized byproduct of the oil extraction industry, usually discarded or used as low-value animal feed through enzyme-aided fractionation, specifically targeting the extraction and conversion of its abundant carbohydrate component, xylan, into emerging prebiotic compounds-xylo-oligosaccharides (XOSs)-which are recognized as promotors of a healthy gut microbiome and overall human wellbeing. An enzymatic treatment using Alcalase® 2.4 L was implemented for facilitating the recovery of a highly pure hemicellulosic fraction (92.2% carbohydrates) rich in ß-(1â4)-linked xylose residues with arabinose and glucuronic acid substitutions (DP-xylan). A further enzymatic treatment of this substrate, using ROHALASE® SEP-VISCO under optimized conditions (70 °C, pH 6, 0.005% v/v enzyme concentration), produced 52.3% of XOSs with a polymerization degree (DP) less than 20 after two hours. Further analyses demonstrated that the majority of the obtained product had a DP less than 6, predominantly consisting of di- and trisaccharides (XOS2 and XOS3) without the significant generation of xylose. These findings highlight the significant potential of SFM for the generation of valuable prebiotic compounds in a sustainable manner.
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This work demonstrates that sesame (Sesamum indicum L.) hull, an unexploited food industrial waste, can be used as an efficient source for the extraction of hemicellulose and/or pectin polysaccharides to further obtain functional oligosaccharides. Different polysaccharides extraction methods were surveyed including alkaline and several enzymatic treatments. Based on the enzymatic release of xylose, arabinose, glucose, and galacturonic acid from sesame hull by using different enzymes, Celluclast®1.5 L, Pectinex®Ultra SP-L, and a combination of them were selected for the enzymatic extraction of polysaccharides at 50 °C, pH 5 up to 24 h. Once the polysaccharides were extracted, Ultraflo®L was selected to produce arabinoxylo-oligosaccharides (AXOS) at 40 °C up to 24 h. Apart from oligosaccharides production from extracted polysaccharides, alternative approaches for obtaining oligosaccharides were also explored. These were based on the analysis of the supernatants resulting from the polysaccharide extraction, alongside a sequential hydrolysis performed with Celluclast®1.5 L and Ultraflo®L of the starting raw sesame hull. The different fractions obtained were comprehensively characterized by determining low molecular weight carbohydrates and monomeric compositions, average Mw and dispersity, and oligosaccharide structure by MALDI-TOF-MS. The results indicated that sesame hull can be a useful source for polysaccharides extraction (pectin and hemicellulose) and derived oligosaccharides, especially AXOS.
Assuntos
Oligossacarídeos , Sesamum , Sesamum/química , Oligossacarídeos/química , Hidrólise , Polissacarídeos/química , Xilanos/química , Xilanos/isolamento & purificação , Pectinas/química , Pectinas/isolamento & purificação , Resíduos Industriais , Arabinose/química , Xilose/químicaRESUMO
BACKGROUND: The general assumption that prebiotics reach the colon without any alterations has been challenged. Some in vitro and in vivo studies have demonstrated that 'non-digestible' oligosaccharides are digested to different degrees depending on their structural composition. In the present study, we compared different methods aiming to assess the digestibility of oligosaccharides synthesized by ß-galactosidase (ß-gal) of Lactobacillus delbruecki subsp. bulgaricus CRL450 (CRL450-ß-gal) from lactose, lactulose and lactitol. RESULTS: In the simulated gastrointestinal fluid method, no changes were observed. However, the oligosaccharides synthesized by CRL450-ß-gal were partially hydrolyzed in vitro, depending on their structure and composition, with rat small intestinal extract (RSIE) and small intestinal brush-border membrane vesicles (BBMV) from pig. Digestion of some oligosaccharides increased when mixtures were fed to C57BL/6 mice used as in vivo model; however, lactulose-oligosaccharides were the most resistant to the physiological conditions of mice. In general ß (1â6) linked products showed higher resistance compared to ß (1â3) oligosaccharides. CONCLUSION: In vitro digestion methods, without disaccharidases, may underestimate the importance of carbohydrates hydrolysis in the small intestine. Although BVMM and RSIE digestion assays are appropriate in vitro methods for these studies, in vivo studies remain the most reliable for understanding what actually happens in the digestion of oligosaccharides. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
Assuntos
Digestão , Camundongos Endogâmicos C57BL , Oligossacarídeos , Prebióticos , beta-Galactosidase , Prebióticos/análise , Animais , beta-Galactosidase/metabolismo , beta-Galactosidase/química , Oligossacarídeos/metabolismo , Oligossacarídeos/química , Camundongos , Ratos , Suínos , Masculino , Lactulose/metabolismo , Lactulose/química , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/química , Intestino Delgado/metabolismo , Intestino Delgado/enzimologia , Lactobacillus/metabolismo , Lactobacillus/enzimologia , Hidrólise , Lactose/metabolismo , Lactose/químicaRESUMO
Inflammatory bowel disease (IBD) is a public health challenge and the use of pectin for symptom amelioration is a promising option. In this work, sunflower pectin has been extracted without (CHP) and with assistance of ultrasound (USP) using sodium citrate as a food-grade extracting agent. At optimal conditions (64 °C, 23 min) the highest yield was obtained with ultrasound application (15.5 vs. 8.1 %). Both pectins were structurally characterized by 1H NMR, HPSEC-ELSD, FT-IR and GC-FID. Unlike CHP, USP showed a lower molecular weight, higher galacturonic acid, lower degree of methyl-esterification and, overall, higher viscosity. These characteristics could affect the anti-inflammatory activity of pectins, evaluated using DSS-induced IBD model mice. So, USP promoted the defence (ICAM-1) and repair of the gastrointestinal mucosa (TFF3, ZO-1) more effectively than CHP. These results demonstrate the potential amelioration of acute colitis in IBD mice through USP supplementation. Taking into account the biomarkers analysed, these results demonstrate, for the first time, the positive impact of sunflower pectin extracted by ultrasound under very soft conditions on inflammatory bowel disease that might open up new possibilities in the treatment of this serious pathology.
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Helianthus , Doenças Inflamatórias Intestinais , Animais , Camundongos , Pectinas/farmacologia , Pectinas/química , Helianthus/química , Espectroscopia de Infravermelho com Transformada de Fourier , Citrato de Sódio , Doenças Inflamatórias Intestinais/diagnóstico por imagem , Doenças Inflamatórias Intestinais/tratamento farmacológicoRESUMO
Arabinoxylan (AX) and arabinoxylo-oligosaccharides (AXOS) derived therefrom are emergent prebiotics with promising health promoting properties, likely linked to its capacity to foster beneficial species in the human gut. Bifidobacteria appear to be one taxa that is frequently promoted following AX or AXOS consumption, and that is known to establish metabolic cross-feeding networks with other beneficial commensal species. Therefore, probiotic bifidobacteria with the capability to metabolize AX-derived prebiotics represent interesting candidates to develop novel probiotic and synbiotic combinations with AX-based prebiotics. In this work we have deepen into the metabolic capabilities of bifidobacteria related to AX and AXOS metabolization through a combination of in silico an in vitro tools. Both approaches revealed that Bifidobacterium longum and, particularly, B. longum subsp. longum, appears as the better equipped to metabolize complex AX substrates, although other related subspecies such as B. longum subsp. infantis, also hold some machinery related to AXOS metabolization. This correlates to the growth profiles exhibited by representative strains of both subspecies in AX or AXOS enriched media. Based on these results, we formulated a differential carbohydrate free medium (CFM) supplemented with a combination of AX and AXOS that enabled to recover a wide diversity of Bifidobacterium species from complex fecal samples, while allowing easy discrimination of AX metabolising strains by the appearance of a precipitation halo. This new media represent an appealing alternative to isolate novel probiotic bifidobacteria, rapidly discriminating their capacity to metabolize structurally complex AX-derived prebiotics. This can be convenient to assist formulation of novel functional foods and supplements, including bifidobacterial species with capacity to metabolize AX-derived prebiotic ingredients.
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Bifidobacterium longum , Simbióticos , Humanos , Bifidobacterium longum/metabolismo , Bifidobacterium/metabolismo , Xilanos , Oligossacarídeos/metabolismo , PrebióticosRESUMO
BACKGROUND: Due to the increasing incidence of obesity and cardiovascular diseases, consumers are demanding products with lower sugar content. In this sense, the reformulation of traditional foods with improved, safe and tasty ingredients is arousing a huge interest. Jams are conventionally produced with elevated amounts of sucrose, which increase the glycaemic index and must be avoided in certain kinds of consumers. RESULTS: This paper describes for the first time the elaboration of strawberry jams using low-methoxyl pectins from sunflower by-products, which allowed the addition of low amounts of sucrose (10-30%). These jams were compared with best-selling commercial samples. An in-depth physicochemical, compositional, sensorial and rheological characterization was carried out. The obtained jams were safe considering aw and pH values; samples presented enough acidity to avoid microorganism development and syneresis. The stabilizing role of sunflower pectin is noteworthy in terms of colour and other physicochemical characteristics. The organoleptic analysis showed that the taste and sweetness of laboratory samples were highly valued, although the presence of pieces of fruits was disliked some panellists. After knowing the content of added sugar used in each jam, the tasters preferred samples with 20% and 30% of sucrose over commercial samples. CONCLUSIONS: The results show the usefulness of sunflower pectin for the elaboration of jams of low glycaemic index. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
Assuntos
Fragaria , Helianthus , Frutas , Helianthus/química , Pectinas/química , SacaroseRESUMO
Worldwide there is great interest in producing low lactose milk and drinks, such as soy beverages, suitable for consumption by lactose-intolerant people. These products have different carbohydrate compositions, which affect quality indicators derived from Maillard reaction (furosine and 5-hydroxyl-methyl furfural, HMF). In this study, quality parameters have been determined in 11 commercial samples (8 lactose-hydrolyzed milks and 3 soy beverages). Presence of ß-galactooligosaccharides in milks (22.2-458.1 mg/100 mL) correlating roughly with the remaining lactose content (1.9-357.7 mg/100 mL). Soy beverages contained α-galactosides in concentrations of 30-75 mg/100 mL. HMF and furosine were detected in all milk samples. In addition, powdered milks subjected to controlled storage (40 °C, aw 0.44) showed a furosine increase up to 88%. In conclusion, a great composition diversity was observed in the different products, probably favored by the lack of regulation and underline the importance of controlling processing and storage conditions to preserve product quality.
RESUMO
Three non-conventional extraction techniques (enzyme-assisted with cellulase, citric acid ultrasound-assisted and enzyme-ultrasound-assisted treatment) and conventional citric acid extraction were applied to obtain pectin from raspberry, blueberry, strawberry and redcurrant, and were compared in terms of extraction yields and physicochemical properties of the extracted pectins. Except for pectin from raspberry, conventional citric acid extraction led to the highest extraction yield (~8%) and, for the same berries, the lowest pectin recovery was found for the extraction with cellulase (~4%). Regarding the structural characteristics of pectins, enzymatically extracted pectins from redcurrant and strawberry exhibited the highest levels of galacturonic acid (≥73%) whereas, in general, this monosaccharide was found from 51 to 69% in the rest of samples. Although, ultrasound-assisted extraction did not improve pectin yield, it minimized the levels of "non-pectic" components leading to the obtainment of purer pectin. The different monomeric composition and the wide range of molecular weight of the obtained pectins pointed out their usefulness in different potential food applications (e.g., thickening, gelling ingredients) and biological activities. This has been evidenced by the differences found in their physicochemical and techno-functional characteristics. Finally, it can be considered that the berries here studied are efficient sources of pectin.
Assuntos
Fracionamento Químico , Frutas/química , Pectinas/isolamento & purificação , Rosales/química , Celulase/química , Ácido Cítrico/química , Fragaria/química , Peso Molecular , Ribes/química , Rubus/química , UltrassomRESUMO
BACKGROUND: This study reports the use of low glycemic sunflower pectin gel, elaborated with calcium and without or with sweeteners (sucrose, stevia and saccharin) as an edible coating and its possible combination with two modified atmosphere packaging (MAP), in order to extend shelf life, maintaining the quality, of strawberries during the storage at 4 °C. RESULTS: This pectin coating, formed with only calcium and/or stevia or saccharin, extended the shelf life of strawberries with respect to uncoating fruits, up to 12 days, keeping the microbial load constant, the firmness and less weight loss. With the same edible coatings, the shelf life of strawberries was extended up to 23 days when they were combined with MAP [10% carbon dioxide (CO2 ), 85% nitrogen (N2 ) and 5% oxygen (O2 )], maintaining the quality of strawberries, while the other MAP, with a higher CO2 concentration (20% CO2 , 75% N2 and 5% O2 ), had no effect. CONCLUSIONS: These results highlight the suitability of the combination of edible pectin coating combined with MAP to obtain an important shelf life extension, maintaining the good quality of the fruit. © 2021 Society of Chemical Industry.
Assuntos
Conservação de Alimentos/métodos , Fragaria/química , Frutas/química , Helianthus/química , Pectinas/química , Extratos Vegetais/química , Filmes Comestíveis , Conservação de Alimentos/instrumentação , Armazenamento de Alimentos , Índice GlicêmicoRESUMO
The intake of diets rich in carbohydrates with a high-glycaemic load provides excessive energy consumption and low nutritional quality, contributing to cardiovascular diseases, type II diabetes and obesity, among other pathologies. Natural options such as the use of low-methoxyl pectin to develop sugar free formulations is growing since they form gel without sucrose, providing stability and viscosity of numerous foodstuffs. In this paper, we have reviewed the consumer habits of sugar intake and the potential of pectin in the elaboration of low-glycaemic index foods, as well as the structure, applications and sources of pectin. Special attention has been paid on the structure-function relationship of low-methoxyl pectins considering their effects on type I and II diabetes. It has been shown that these pectins reduce the post-prandial glycaemic responses and have an important and recognised potential for the treatment and prevention of diabetes. Rheological behaviour of pectin, impact on intestinal microbiota and on different biomarkers have been postulated as the potential involved mechanisms. As future trends, it is necessary to consider new sources and structures of pectin that present improved functionalities. For this purpose, the obtainment of pectin and derivatives should be placed in a framework of sustainable food systems.
Assuntos
Diabetes Mellitus Tipo 2 , Pectinas , Dieta , Humanos , Reologia , ViscosidadeRESUMO
Holder pasteurization (HoP; 62.5 °C, 30 min) is commonly used to ensure the microbiological safety of donor human milk (DHM) but diminishes its nutritional properties. A high-temperature short-time (HTST) system was designed as an alternative for human milk banks. The objective of this study was to evaluate the effect of this HTST system on different nutrients and the bile salt stimulated lipase (BSSL) activity of DHM. DHM was processed in the HTST system and by standard HoP. Macronutrients were measured with a mid-infrared analyzer. Lactose, glucose, myo-inositol, vitamins and lipids were assayed using chromatographic techniques. BSSL activity was determined using a kit. The duration of HTST treatment had a greater influence on the nutrient composition of DHM than did the tested temperature. The lactose concentration and the percentage of phospholipids and PUFAs were higher in HTST-treated than in raw DHM, while the fat concentration and the percentage of monoacylglycerides and SFAs were lower. Other nutrients did not change after HTST processing. The retained BSSL activity was higher after short HTST treatment than that following HoP. Overall, HTST treatment resulted in better preservation of the nutritional quality of DHM than HoP because relevant thermosensitive components (phospholipids, PUFAs, and BSSL) were less affected.
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The aim of this work was to develop a comprehensive workflow to elucidate molecular features of artichoke pectic oligosaccharides (POS) contributing to high potential prebiotic activity. First, obtainment of artichoke POS by Pectinex® Ultra-Olio was optimised using an artificial neural network. Under optimal conditions (pH 6.86; 1.5 h; enzyme dose 520.5 U/g pectin) POS yield was 624 mg/g pectin. Oligosaccharide structures (Mw < 1.3 kDa) were characterised by MALDI-TOF-MS. Then, conformational analysis of glycosidic bonds was performed by replica exchange molecular dynamics simulations and interaction mechanisms between POS and several microbial glycosidases were proposed by molecular modelling. Chemical information was integrated in virtual simulations of colonic fermentation. Highest hydrolysis rate was obtained for GalA-Rha-GalA trisaccharide, while the presence of partial negative charges and high radius of gyration enhance short chain fatty acid formation in distal colon. Established structure-activity relationships could help the rational design of prebiotics and clinical trials.
Assuntos
Colo/metabolismo , Cynara scolymus/química , Ácidos Graxos Voláteis/biossíntese , Oligossacarídeos/metabolismo , Pectinas/metabolismo , Prebióticos/análise , Análise Fatorial , Ácidos Graxos Voláteis/química , Fermentação , Concentração de Íons de Hidrogênio , Hidrólise , Simulação de Dinâmica Molecular , Redes Neurais de Computação , Oligossacarídeos/química , Oligossacarídeos/isolamento & purificação , Pectinas/química , Pectinas/isolamento & purificação , Extratos Vegetais/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Relação Estrutura-Atividade , Interface Usuário-ComputadorRESUMO
Galactooligosaccharides (GOS) are useful dietary ingredients recognized worldwide as prebiotics. In the present study, we evaluated the ß-galactosidase (ß-gal) activity of a panel of lactic acid bacteria (LAB) in order to select strains for the synthesis of oligosaccharides from lactose (GOS) and lactulose (OsLu) with a potential prebiotic effect. Fifteen strains out of 20 were able to grow on lactose and showed ß-gal activities between 0.03 and 2.06 U mg-1, whereas eleven were able to synthesize GOS. Lactobacillus delbrueckii subsp. bulgaricus CRL450, the strain with the highest ß-gal activity, synthesized a maximum of 41.3% GOS and 21.0% OsLu from lactose and lactulose, respectively, with ß-(1 â 6) and secondary ß-(1 â 3) linkages. When these compounds were tested without purifying, as carbon sources for the development of recognized probiotics and the producer strain, high growth was observed compared to non-prebiotic sugars like glucose and lactose. When the purified oligosaccharides were tested, the bacterial growth decreased, but the microorganisms displayed metabolic activity evidenced by the consumption of carbohydrates and the production of lactic acid. Additionally, the purified oligosaccharides demonstrated a bifidogenic effect. The obtained results support the potential of L. delbrueckii subsp. bulgaricus CRL450 for the production of the prebiotics GOS and OsLu and encourage the optimization of their synthesis for the design of new functional food ingredients.
Assuntos
Fermentação , Galactose/metabolismo , Lactobacillus delbrueckii/metabolismo , Lactose/metabolismo , Lactulose/metabolismo , Oligossacarídeos/biossíntese , Prebióticos , Humanos , Probióticos , beta-Galactosidase/metabolismoRESUMO
The degree of digestion, modulated by rat small intestinal extract on different functional fibers was investigated. In general, inulin-type fructans and fructooligosaccharides were the most resistant to the enzymatic digestion. Results evidenced the high-resistance of fructosyl-fructose bonds. This fits well with the concept of prebiotic carbohydrates. However, the mixture of melibiose, manninotriose and verbascotetraose (α-GOS) from peas, with a considerably lower molecular weight (0.6 kDa) than the fructans studied, were highly digested (61.2%). Interestingly, the Gal-(1 â 6)-Gal bonds present into the manninotriose and verbascotetraose were more prone to be hydrolyzed than Gal-(1 â 6)-Glc (melibiose). However, when melibiose was the only disaccharide present in the reaction mixture, the hydrolysis was also high (67.7%). The use of small intestinal enzymatic preparations is a realistic approximation to evaluate the digestion of different carbohydrates, thus, showing that recognized non-digestible carbohydrates can also be partially digested.
Assuntos
Metabolismo dos Carboidratos , Enzimas/metabolismo , Intestino Delgado/química , Polissacarídeos/química , Extratos de Tecidos/farmacologia , Animais , Carboidratos/química , Digestão , Prebióticos , Ratos , Extratos de Tecidos/química , Extratos de Tecidos/metabolismoRESUMO
It is well known that one of the main problems in galactooligosaccharide production (GOS) via tranglycosylation of lactose is the presence of monosaccharides that contribute to increasing the glycaemic index, as is the case of glucose. In this work, as well as studying the effect of ultrasound (US) on glucose oxidase (Gox) activation during gluconic acid (GA) production, we have carried out an investigation into the selective oxidation of glucose to gluconic acid in multienzymatic reactions (ß-galactosidase (ß-gal) and Gox) assisted by power US using different sources of lactose as substrate (lactose solution, whey permeate, cheese whey). In terms of the influence of matrix on GOS and GA production, lactose solution gave the best results, followed by cheese whey and whey permeate, salt composition being the most influential factor. The highest yields of GOS production with the lowest glucose concentration and highest GA production were obtained with lactose solution in multienzymatic systems in the presence of ultrasound (30% amplitude) when Gox was added after 1â¯h of treatment with ß-gal. This work demonstrates the ability of US to enhance efficiently the obtainment of prebiotic mixtures of low glycaemic index.
Assuntos
Enzimas/metabolismo , Galactose/metabolismo , Gluconatos/metabolismo , Lactose/metabolismo , Oligossacarídeos/metabolismo , Sonicação , Soro do Leite/química , PrebióticosRESUMO
A pilot-scale extraction of sunflower pectin with 0.74% (w/v) sodium citrate (72 °C, 194 min) and different procedures of purification including alcohol precipitation, ultrafiltration (UFDF) and microfiltration (MFDF) with diafiltration were carried out. Considering the alcohol treatment, the yields were similar at laboratory and pilot-scale (~8.9%), demonstrating the efficiency of the scale-up. With respect to membrane processes, the best results were obtained with UFDF, showing the highest yield (13.3%) and pectin concentration higher than 90%. In all cases, pectins presented very low amount (~1%) of glucose and mannose, monosaccharides not included in the pectin structure. Detailed NMR analysis and functional properties (emulsifying and viscosity) that were also assessed corroborated the good quality of UFDF obtained pectin. These results point out that the obtainment of sunflower pectin of good quality can be achieved at pilot-scale by the extraction with sodium citrate and purification with membrane separation, eco-friendly alternatives to conventional procedures.
Assuntos
Helianthus/química , Pectinas/química , Pectinas/isolamento & purificação , Fracionamento Químico , Emulsificantes/química , Citrato de Sódio/química , Ultrafiltração/métodos , ViscosidadeRESUMO
Anti-inflammatory properties of artichoke pectin and modified fractions (arabinose- and galactose-free) used at two doses (40 and 80 mg kg-1) in mice with colitis induced by dextran sulfate sodium have been investigated. Expression of pro-inflammatory markers TNF-α and ICAM-I decreased in groups of mice treated with original and arabinose-free artichoke pectin while IL-1ß and IL-6 liberation was reduced only in mice groups treated with original artichoke pectin. A decrease in iNOS and TLR-4 expression was observed for most treatments. Intestinal barrier gene expression was also determined. MUC-1 and Occludin increased in groups treated with original artichoke pectin while MUC-3 expression also increased in arabinose-free pectin treatment. Galactose elimination led to a loss of pectin bioactivity. Characteristic expression profiles were established for each treatment through artificial neural networks showing high accuracy rates (≥90%). These results highlight the potential amelioration of inflammatory bowel disease on mice model colitis through artichoke pectin administration.
Assuntos
Anti-Inflamatórios/administração & dosagem , Colite/tratamento farmacológico , Cynara scolymus/química , Pectinas/administração & dosagem , Extratos Vegetais/administração & dosagem , Animais , Colite/induzido quimicamente , Colite/genética , Colite/imunologia , Sulfato de Dextrana/efeitos adversos , Modelos Animais de Doenças , Humanos , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Intestinos/efeitos dos fármacos , Intestinos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologiaRESUMO
Pectic oligosaccharides (POS) from citrus and apple pectin hydrolysis using ViscozymeL and Glucanex200G have been obtained. According to the results, maximum POS formation was achieved from citrus pectin after 30 min of hydrolysis with ViscozymeL, with a yield of 652 mg g-1 and average molecular mass ( Mw) of 0.8-2.5 kDa, while with Glucanex200G, the yield was 518 mg g-1 and Mw was 0.8-7.1 kDa. Digalacturonic and trigalacturonic acids were identified among other low Mw compounds as di- and tri-POS. In addition, differences in GC-MS spectra of all oligosaccharides found in the hydrolysates were studied by employing random forests and other algorithms to identify structural differences between the obtained POS, and high prediction rates were shown for new samples. Chemical structures were proposed for some influential m/ z ions, and 12 association rules that explain differences according to pectin and enzyme origin were built. This information could be used to establish structure-function relationships of POS.
Assuntos
Citrus/química , Malus/química , Complexos Multienzimáticos/química , Oligossacarídeos/química , Pectinas/química , Extratos Vegetais/química , Biocatálise , Cromatografia Gasosa-Espectrometria de Massas , Hidrólise , Peso MolecularRESUMO
Lactulose-derived oligosaccharides (OsLu) are prebiotic galactooligosaccharides (GOS) beneficial for human health including immunomodulatory properties; however, the molecular mechanism is unclear. OsLu produced by enzymatic synthesis can be purified with Saccharomyces cerevisiae (OsLu-Sc). We show that this purification introduces yeast-derived proteins reactive to Dectin-2, an innate immune receptor for fungal polysaccharides. Using a cell-based bioassay, we tested the binding of OsLu and GOS samples to Dectin-2. While OsLu purified with active charcoal and commercial GOS failed to bind to Dectin-2, we found OsLu-Sc bound to this receptor. The carbohydrate-binding incompetent mutant of Dectin-2 failed to bind to OsLu-Sc. These data suggest that OsLu-Sc introduced carbohydrate ligands for Dectin-2. In accordance with this, proteomic analysis revealed OsLu-Sc contained S. cerevisiae-derived mannoproteins. Therefore, our data highlight the importance of the purification method for OsLu, which may positively affect the bioactivity of OsLu. Data are available via ProteomeXchange with identifier PXD010495.
Assuntos
Proteínas Fúngicas/metabolismo , Lactulose/química , Lectinas Tipo C/metabolismo , Oligossacarídeos/análise , Receptores Imunológicos/metabolismo , Saccharomyces cerevisiae/metabolismo , Prebióticos , ProteômicaRESUMO
The potential of probiotic bacteria to produce prebiotic oligosaccharides by transgalactosylation has been minimally studied. In this work, we screened the ß-galactosidase (ß-gal) activity of dairy propionibacteria (PAB) isolated from Argentinean foods to select strains for the synthesis of oligosaccharides from lactose (GOS) and lactulose (OsLu). PAB, when grown in a medium with lactose as a carbon source, were disrupted, and the cell-free extracts were assayed for ß-gal activity. Nine strains grew on lactose and showed ß-gal activities from 0.27 to 2.60 U mL-1. Propionibacterium acidipropionici LET 120, the strain with the highest activity, was able to synthesize, using 30% lactose and lactulose at pH 6.5 and 45⯰C, 26.8% of LET 120-GOS and 26.1% of LET 120-OsLu after 24â¯h. When they were tested as carbon sources for growth, P. acidipropionici LET 120 attained higher biomasses, µmax and ß-gal activities at the expense of Aspergillus oryzae-OsLu, Vivinal®-GOS and lactulose compared to lactose or glucose. In addition, LET 120-GOS and LET 120-OsLu synthesized by PAB were prebiotic for some probiotic strains. For the first time, our results show the production of GOS and OsLu by dairy PAB, and these results encourage further studies on the optimization of the synthesis and structure characterization of the obtained oligosaccharides.