Low Molecular Weight Oligosaccharides Research Articles

Cyanometabolites: molecules with immense antiviral potential

Cyanometabolites are active compounds derived from cyanobacteria that include small low molecular weight peptides, oligosaccharides, lectins, phenols, fatty acids, and alkaloids. Some of these compounds may pose a threat to human and environment. However, majority of them are known to have various health benefits with antiviral properties against pathogenic viruses including Human immunodeficiency virus (HIV), Ebola virus (EBOV), Herpes simplex virus (HSV), Influenza A virus (IAV) etc. Cyanometabolites classified as lectins include scytovirin (SVN), Oscillatoria agardhii agglutinin (OAAH), cyanovirin-N (CV-N), Microcystis viridis lectin (MVL), and microvirin (MVN) also possess a potent antiviral activity against viral diseases with unique properties to recognize different viral epitopes. Studies showed that a small linear peptide, microginin FR1, isolated from a water bloom of Microcystis species, inhibits angiotensin-converting enzyme (ACE), making it useful for the treatment of coronavirus disease 2019 (COVID-19). Our review provides an overview of the antiviral properties of cyanobacteria from the late 90s till now and emphasizes the significance of their metabolites in combating viral diseases, particularly severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which has received limited attention in previous publications. The enormous medicinal potential of cyanobacteria is also emphasized in this review, which justifies their use as a dietary supplement to fend off pandemics in future.

Hyaluronan and Reactive Oxygen Species Signaling-Novel Cues from the Matrix?

Hyaluronan (HA) is a naturally occurring non-sulfated glycosaminoglycan (GAG) localized to the cell surface and the tissue extracellular matrix (ECM). It is composed of disaccharides containing glucuronic acid and N-acetylglucosamine, is synthesized by the HA synthase (HAS) enzymes and is degraded by hyaluronidase (HYAL) or reactive oxygen and nitrogen species (ROS/RNS) actions. HA is deposited as a high molecular weight (HMW) polymer and degraded to low molecular weight (LMW) fragments and oligosaccharides. HA affects biological functions by interacting with HA-binding proteins (hyaladherins). HMW HA is anti-inflammatory, immunosuppressive, and antiangiogenic, whereas LMW HA has pro-inflammatory, pro-angiogenetic, and oncogenic effects. ROS/RNS naturally degrade HMW HA, albeit at enhanced levels during tissue injury and inflammatory processes. Thus, the degradation of endothelial glycocalyx HA by increased ROS challenges vascular integrity and can initiate several disease progressions. Conversely, HA exerts a vital role in wound healing through ROS-mediated HA modifications, which affect the innate immune system. The normal turnover of HA protects against matrix rigidification. Insufficient turnover leads to increased tissue rigidity, leading to tissue dysfunction. Both endogenous and exogenous HMW HA have a scavenging capacity against ROS. The interactions of ROS/RNS with HA are more complex than presently perceived and present an important research topic.

Pectooligosaccharides rich in galacturonic acid produced from Orange Processing Waste by autohydrolysis: Process optimization and kinetic analysis

Pectooligosaccharides (POS) are notorious for their prebiotics and elicitor activities. In this work, we have optimized the autohydrolysis of Orange Processing Waste (OPW) to obtain POS fractions via a Taguchi approach, maximizing homogalacturonan oligosaccharides yield by considering several process variables: particle diameter, stirring speed, liquid-solid ratio and time, determining that only time had a significant effect in the studied range. 19.38 % w/w POS yield from OPW was obtained in optima conditions. After ensuring no limitation due to mass transfer, the kinetic analysis assumed that homogalacturonan was converted into high, medium and low molecular weight oligosaccharides, and galacturonic acid. Moreover, we considered that there is one pseudo-homogeneous reaction type combined with three irreversible, elemental, of pseudo first global order (first order with respect to the reagent which is hydrolysed) reactions in series. The model successfully reproduced experimental data at constant temperature (150 °C–180 °C) and pH (3.73).

Mechanocatalytic depolymerization of hemicellulose to low molecular weight oligosaccharides over an aquivion ionomer

Hemicellulose is mechanocatalytically depolymerized to low molecular weight oligosaccharides with a DP lower than 9 in 70% yield, and without any solvent.

Low-molecular weight oligosaccharides from gum tragacanth (Astragalus gossypinus) ameliorate nonalcoholic fatty liver disease (NAFLD) in Wistar male rats.

Overall, the results of the current study demonstrated that the use of GT oligosaccharides obtained from Gum tragacanth (Astragalus gossypinus) showed significant antioxidant properties and hypoglycemia in NAFLD induced rats and could be used as a useful nutritional strategy for the prevention and treatment of NAFLD.

Anti-SARS-CoV-2 and anticoagulant properties of Pentacta pygmaea fucosylated chondroitin sulfate depend on high molecular weight structures.

Fucosylated chondroitin sulfate (FucCS) is a unique marine glycosaminoglycan that exhibits diverse biological functions, including antiviral and anticoagulant activity. In previous work, the FucCS derived from Pentacta pygmaea (PpFucCS) showed moderate anticoagulant effect but high inhibitory activity against the Wuhan strain of severe acute respiratory syndrome coronavirus (SARS-CoV-2). In this study, we perform free-radical depolymerization of PpFucCS by the copper-based Fenton method to generate low molecular weight (MW) oligosaccharides. PpFucCS oligosaccharides were structurally analyzed by 1H nuclear magnetic resonance spectroscopy and were used to conduct structure-activity relationship studies regarding their effects against SARS-CoV-2 and clotting. Anticoagulant properties were measured by activated partial thromboplastin time, protease (factors Xa and IIa) inhibition by serine protease inhibitors (antithrombin [AT] and heparin cofactor II [HCII]), and competitive surface plasmon resonance (SPR) assay using AT, HCII, and IIa. Anti-SARS-CoV-2 properties were measured by the concentration-response inhibitory curves of HEK-293T-human angiotensin-converting enzyme-2 cells infected with a baculovirus pseudotyped SARS-CoV-2 Delta variant spike (S)-protein and competitive SPR assays using multiple S-proteins (Wuhan, N501Y [Alpha], K417T/E484K/N501Y [Gamma], L542R [Delta], and Omicron [BA.2 subvariant]). Cytotoxicity of native PpFucCS and oligosaccharides was also assessed. The PpFucCS-derived oligosaccharide fraction of the highest MW showed great anti-SARS-CoV-2 Delta activity and reduced anticoagulant properties. Results have indicated no cytotoxicity and MW dependency on both anti-SARS-CoV-2 and anticoagulant effects of PpFucCS, as both actions were reduced accordingly to the MW decrease of PpFucCS. Our results demonstrate that the high-MW structures of PpFucCS is a key structural element to achieve the maximal anti-SARS-CoV-2 and anticoagulant effects.

Effect of drying method and process conditions on physicochemical and rheological properties of arabinoxylans extracted from corn-lime-cooking-liquor on a pilot plant scale

Corn-lime-cooking-liquor (nejayote), which is a pollutant wastewater from the corn-tortilla-industry, contains valuable products, such as arabinoxylans (AXs) and phenolics. Recently, an innovative process that employs enzymatic and ultrafiltration processes for recovering and purifying AXs from nejayote was described, although neither the drying methods nor the physicochemical and rheological properties of the recovered AXs by this method at the pilot plant scale have been studied. Nejayote from a batch artisanal process (NBAP) and from a continuous industrial process (NCIP) were compared with a corn-bran-hydrolysate (CBH) as sources for AXs obtention. Freeze drying (FD), spray drying (SD) and solvent drying (SoD) were assessed as processes for drying purified and concentrated AXs solutions. Regardless of the process, AXs obtained from nejayote contain 3 times less (0.30 mg/g) esterified ferulic acid (EFA) than did AXs from corn bran (1.01 mg/g) and thus fail to yield elastic gels upon oxidative cross-linking. Molecular weight distribution (MwD) and dynamic viscosity data show that AXs from nejayote are shorter than AXs from corn bran. In general, the drying method had no effect on the physicochemical and rheological properties of AXs, although additional purification using ethanol precipitation and SoD, eliminated low molecular weight oligosaccharides and residual salts resulting from the neutralization of the corn-lime-cooking-liquor. Spray drying seems to be the most cost-effective unit operation for the industrial production of AXs from nejayote without affecting their physicochemical characteristics.

Low Molecular Weight Oligosaccharide from Panax ginseng C.A. Meyer against UV-Mediated Apoptosis and Inhibits Tyrosinase Activity In Vitro and In Vivo.

To find new anti-UV and whitening agents, 21 fractions isolated from three preparations of ginseng (white, red, and black ginseng) were screened, and their antioxidant effects on AAPH- or H2O2-induced damage were investigated. Furthermore, the protective effect against UV-mediated apoptosis and the tyrosinase inhibitory activity of the targeted fractions were evaluated in vitro and in a zebrafish model. Among all fractions, F10 from white ginseng was selected as having the strongest anti-UV and antimelanogenesis activities. This fraction exhibited excellent inhibitory effects on the pigmentation of zebrafish, which may be due to its potential tyrosinase inhibitory activity. Additionally, the chemical composition of F10 was evaluated by UPLC-MS and NMR instruments. The results indicated that F10 had a carbohydrate content of more than 76%, and the weight-average molecular weight was approximately 239 Da. Disaccharide sucrose was the main active compound in F10. These results suggest that F10 could be used as an ingredient for whitening cosmetics and regarded as an anti-UV filter in the future.

Streptococcus infantarius 25124 isolated from pozol produces a high molecular weight amylopullulanase, a key enzyme for niche colonization

Abstract Pozol is a beverage made with maize dough that is prepared after boiling the kernels in limewater, causing a decrease in soluble sugars, with starch being the main fermentable carbohydrate in the dough. Previously, Streptococcus infantarius ssp. infantarius 25124 (Sii-25124) was identified as the most amylolytic bacteria isolated in this product. Analysis of Sii-25124 amylolytic enzymes revealed two amylases, a cytoplasmic α-amylase of 55.7 kDa and an extracellular amylopullulanase of 246.3 kDa, with two catalytic domains, one typical of an α-amylase and another typical of a pullulanase/glycogen debranching enzyme. Characterization of the joint activity of both enzymes using Sii-25124 cell lysate supernatant demonstrated stability between 30 °C and 45°C, and pH stability in a range between 6.8 and 8.0. The joint activity of Sii-25124 amylases showed a fast production of reducing sugars when starch was used as the substrate. In contrast, reducing sugar production from amylopectin was lower, but it steadily increased throughout the reaction time. The amylopullulanase produced by Sii-25124 hydrolyzes the starch in the dough to produce low molecular weight oligosaccharides, which may be transported into Sii-25124 cells, so that intracellular α-amylase hydrolyzes them to mono- and disaccharides. Amylopullulanase production by Sii-25124 could be an example of a specialized enzyme that successfully dominates starchy food fermentation.

Gentianose: Purification and structural determination of an unknown oligosaccharide in grape seeds

Grape seeds are among the main constituents of grape pomace, ranging between 20% and 30% of the wet matrix; however, their oligosaccharide composition has not been studied. This paper describes the purification and the identification of low molecular weight oligosaccharides contained in an EtOH/water extract of grape seeds. A sequential two-step purification by size exclusion chromatography was carried out to fractionate compounds according to molecular weights. Chemical characterization of the combined fractions was performed by Magnetic Resonance Spectroscopy and Gas Chromatography-Mass Spectrometry analyses. The separation process gave two fractions abundant in sucrose and glucose. A third fraction containing trisaccharides was acetylated allowing the purification of the main trisaccharide. The structure elucidation of the acetylated product made it possible to identify gentianose, a predominant carbohydrate reserve found in the storage roots of perennial Gentiana lutea. Grape seeds are wine industry by-products and the obtained results suggest the importance of their recovery.

Characterization of a bifunctional alginate lyase as a new member of the polysaccharide lyase family 17 from a marine strain BP-2

ObjectivesBifunctional alginate lyase can efficiently saccharify alginate biomass and prepare functional oligosaccharides of alginate.ResultsA new BP-2 strain that produces alginate lyase was screened and identified from rotted Sargassum. A new alginate lyase, Alg17B, belonging to the polysaccharide lyase family 17, was isolated and purified from BP-2 fermentation broth by freeze-drying, dialysis, and ion exchange chromatography. The enzymatic properties of the purified lyase were investigated. The molecular weight of Alg17B was approximately 77 kDa, its optimum reaction temperature was 40–45 °C, and its optimum reaction pH was 7.5–8.0. The enzyme was relatively stable at pH 7.0–8.0, with a temperature range of 25–35 °C, and the specific activity of the purified enzyme reached 4036 U/mg. A low Na+ concentration stimulated Alg17B enzyme activity, but Ca2+, Zn2+, and other metal ions inhibited it. Substrate specificity analysis, thin-layer chromatography, and mass spectrometry showed that Alg17B is an alginate lyase that catalyses the hydrolysis of sodium alginate, polymannuronic acid (polyM) and polyguluronic acid to produce monosaccharides and low molecular weight oligosaccharides. Alg17B is also bifunctional, exhibiting both endolytic and exolytic activities toward alginate, and has a wide substrate utilization range with a preference for polyM.ConclusionsAlg17B can be used to saccharify the main carbohydrate, alginate, in the ethanolic production of brown algae fuel as well as in preparing and researching oligosaccharides.

Applications of Algal Polysaccharides and Derivatives in Therapeutic and Agricultural Fields.

Recently, researchers have given more and more consideration to natural polysaccharides thanks to their huge properties such as stability, biodegradability and biocompatibility for food and therapeutics applications. a number of enzymatic and chemical processes were performed to generate bioactive molecules, such as low molecular weight fractions and oligosaccharides derivatives from algal polysaccharides. These considerable characteristics allow algal polysaccharides and their derivatives such as low molecular weight polymers and oligosaccharides structures to have great potential to be used in lots of domains, such as pharmaceutics and agriculture etc. Conclusion: The present review describes the mains polysaccharides structures from Algae and focuses on the currents agricultural (fertilizer, bio-elicitor, stimulators, signaling molecules and activators) and pharmaceutical (wound dressing, tissues engineering and drugs delivery) applications by using polysaccharides and/or their oligosaccharides derivatives obtained by chemical, physical and enzymatic processes.

Enzymatic treatments as alternative to produce chitin fragments of low molecular weight from Alternaria alternata

ABSTRACTChitin and its oligosaccharides are involved in the plant defense response against fungal pathogens. In most studies, these molecules come from crustacean shell, and there are scarce studies on the use of fungal chitin. Usually the extraction of chitin is by alkaline treatments, which affect the acetylation degree (DA), and the obtaining of oligosaccharides of low molecular weight; so the use of enzymes is proposed as an alternative treatment to obtain chitin oligosaccharides from fungi. The objective of this work was to characterize the chitin fragments of Alternaria alternata extracted by alkaline and enzymatic treatments. The chitin extracted was ultrasonicated and ultrafiltrated to produce chitin fragments. Enzymatic treatments decreased the protein content to 29%, and the bands corresponding to β‐glucans decreased when β‐1,3‐glucanase was applied. Fragments smaller than 1 kDa, DA of 76.7%, and 35.4 μg glucosamine/mg dry weight were obtained by enzymatic treatments. The enzymatic treatments show promising results to extract chitin fragments from A. alternata without severely affect the DA of the molecule. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 47339.

Stabilizing effect of fos during freeze-drying and storage of lactobacillus delbrueckiI SUBSP. Bulgaricus results from a balance between high and low molecular weight oligosaccharides

Stabilizing effect of fos during freeze-drying and storage of lactobacillus delbrueckiI SUBSP. Bulgaricus results from a balance between high and low molecular weight oligosaccharides

The alginate lyase from Isoptericola halotolerans CGMCC 5336 as a new tool for the production of alginate oligosaccharides with guluronic acid as reducing end

Alginate oligosaccharides are depolymerization products of alginate by enzymatic degradation or physicochemical treatments. Alginate lyase has received great interests due to its use in oligosaccharide preparation. The substrate specificity of alginate lyase directly determines the type and degree of polymerization of alginate oligosaccharides. In this paper, the degradation products of Alginate lyase from Isoptericola halotolerans CGMCC 5336 were size-fractionated by gel filtration chromatography and the fractions were characterized by TLC, HPLC, MS and 1H NMR spectroscopy. The Mw values for these fractions were 352, 528, 704 Da which represented di-, tri-, and tetrasaccharide fragments, respectively. The 1H NMR revealed that the non-reducing ends of these oligosaccharides were 4-deoxy-L-erythro-hex-4-enepyranosyluronate and the reducing ends of these oligosaccharides were all guluronic acid. Therefore, it is believed that the alginate lyase produced by Isoptericola halotolerans CGMCC 5336 was capable of performing β elimination on guluronic acid residue so that low molecular weight oligosaccharides with guluronic acid on the reducing end could be obtained in this way.

Structural, functional, and biological properties of potato peel oligosaccharides

Enzymatic hydrolysis of water-soluble polysaccharides from potato peel waste (PPPW) generates low molecular weight oligosaccharides with a yield of 63%. The oligosaccharides generated from potato peel polysaccharides (OPPP) were purified by Superdex-30 column. The results showed the presence of 8 peaks (OPPP1–OPPP8). The identification of all the fractions by chromatography analysis (GC-FID) illustrated that the most prominent residues were glucose with traces of galactose, arabinose and rhamnose. Finally, mass spectrometry (MALDI-ToF-ToF) analysis showed that the generated oligosaccharides were heterogeneous and contained different degree of polymerization (DP). Indeed, the obtained oligosaccharides fractions OPPP3, OPPP4, OPPP5, OPPP6 and OPPP7 were compose of the following degree of polymerization DP5; DP4; DP2; DP1 and DP1, respectively. Potato peel oligosaccharides (OPPP) efficiency were tested using different concentrations in functional properties. The results showed good foaming and emulsion properties. This study also aimed to investigate the antioxidant activity of OPPP. The items explored included the DPPH radical-scavenging capacity (IC50 OPPP=2.5mg/mL), reducing power (OD: 0.622±0.032 at a concentration of 20mg/mL), β-carotene bleaching inhibition activity (45.335±3.653%), and also the ABTS radical scavenging activity (14.835±0.1%).These findings indicate that potato peel oligosaccharides have potent antioxidant activities. Hence, one can suggest that these oligosaccharides might contribute as additives in food, pharmaceutical and cosmetic preparations.

Carrageenans and carrageenases: versatile polysaccharides and promising marine enzymes

Carrageenans are sulfated polysaccharides isolated from marine red algae that share a common backbone of D-galactose alternately linked by α(1,3) and β(1,4) glycosidic linkages. They are classified based on the number and the position of the sulfate ester groups and the occurrence of a 3,6 anhydro-ring in the α-linked galactose. Accordingly the three most commercially exploited carrageenans are κ-, ι-, and λ-carrageenans. Because of their biocompatibility, exceptional physicochemical features and emulsifying, thickening, gelling and stabilizing abilities, they have found several industrial application, especially in food, pharmaceutical and cosmetic industries. Moreover, carrageenans can be degraded into lower molecular weight oligosaccharides, which have been reported to have promising pharmacological properties and potential therapeutic applications. Enzymes which degrade carrageenans are called carrageenases and are produced only by marine bacterial species. These enzymes all are endohydrolases that hydrolyze the internal β 1,4 linkages in carrageenans and produce a series of homologous even-numbered oligosaccharides with various biological and physiological activities including anti-tumor, anti-inflammation, anti-viral, anti-coagulation, etc. Carrageenase enzymes have also other applications related to the biomedical field, bioethanol production, prevention of red algal bloom, obtaining algal protoplasts, etc. In the first part of this review, general information regarding structure, physicochemical properties, biological activities and potential applications of carrageenans is summarized. The second part deals with research and development works on some aspects of carrageenase enzymes like the source, characterization, the kinetics and biochemical properties and their applications in various industries.

Prebiotic Oligosaccharides: Special Focus on Fructooligosaccharides, Its Biosynthesis and Bioactivity.

The bacterial groups in the gut ecosystem play key role in the maintenance of host's metabolic and structural functionality. The gut microbiota enhances digestion processing, helps in digestion of complex substances, synthesizes beneficial bioactive compounds, enhances bioavailability of minerals, impedes growth of pathogenic microbes, and prevents various diseases. It is, therefore, desirable to have an adequate intake of prebiotic biomolecules, which promote favorable modulation of intestinal microflora. Prebiotics are non-digestible and chemically stable structures that significantly enhance growth and functionality of gut microflora. The non-digestible carbohydrate, mainly oligosaccharides, covers a major part of total available prebiotics as dietary additives. The review describes the types of prebiotic low molecular weight carbohydrates, i.e., oligosaccharides, their structure, biosynthesis, functionality, and applications, with a special focus given to fructooligosaccharides (FOSs). The review provides an update on enzymes executing hydrolytic and fructosyltransferase activities producing prebiotic FOS biomolecules, and future perspectives.

Thermostable multifunctional GH74 xyloglucanase from Myceliophthora thermophila: high-level expression in Pichia pastoris and characterization of the recombinant protein.

A xyloglucanase of the GH74 family was identified in the thermophilic fungus strain Myceliophthora thermophila VKPM F-244, and its gene sequence was optimized for cloning and expression in Pichia pastoris. The recombinant xyloglucanase MtXgh74 exhibited the highest activity toward tamarind seed xyloglucan with a K M value of 0.51±0.06mg/mL. The activities on barley β-glucan and carboxymethylcellulose were about 4 and 2%, respectively, compared to xyloglucan. Maximum xyloglucanase activity was observed at 70-75°C and pH6.5. After pre-incubation at 50°C, pH6.0 for 3h, the enzyme retained 100% of its activity. The half-life of MtXgh74 at 60°C, pH6.0 was 40min. In P. pastoris, MtXgh74 was produced in glycosylated form. The enzyme production in a 1 L bioreactor resulted in a yield of 118U/mL or 5.3g/L after 51 h fermentation. Kinetic studies of the hydrolysis product formation suggest that MtXgh74 has an endo-processive mode of action. The final products were the standard xyloglucan building blocks XXXG, XXLG, XLXG, and XLLG. Additionally, MtXgh74 hydrolyzed various linkages within the xyloglucan building blocks XXXG, XXLG, and XLXG (except XLLG) producing diverse low molecular weight oligosaccharides which may be identified by MALDI-TOF as XG, XX, XXG/GXX/XGX, XXX, LG, LX/XL, XLX/XXL, LLG, GXXXG, GXLLG, XLLGX. The unique combination of different activities within one enzyme along with its high thermostability and specificity toward xyloglucan makes MtXgh74 a promising candidate enzyme for industrial applications.

Influence of DE-value on the physicochemical properties of maltodextrin for melt extrusion processes

In this study, five different types of maltodextrins (DE-2, DE-6, DE-12, DE-17 and DE-19) were characterized for the physico-chemical properties. TGA, DVS and SEC analyses were carried out and additionally apparent melt-viscosity (in a micro-extruder) and the glass transition temperature (analyzed by DMA) of maltodextrin/plasticizer mixtures were also measured in order to evaluate both the effect of plasticizer nature and content and the effect of the DE-value. For this, three plasticizing agents were compared: water, d-sorbitol and glycerin. The adsorption isotherms showed that depending on the DE-value and the relative humidity they were exposed to, different behavior could be obtained. For example, for relative humidities below 60% RH maltodextrin DE-2 was the least hygroscopic. And on the contrary for relative humidities above 75% RH maltodextrin DE-2 was the most hygroscopic. The rheology measurements showed that the viscosity decreased with the increase of the DE-value and with the plasticizer content, as expected. On the contrary, no direct correlation could be established between the DE-value and the glass transition temperature. These results demonstrated that to predict maltodextrins behavior and to better adapt the process conditions, combined analyses are mandatory as the DE-value alone is not sufficient. The most compelling evidence was obtained by size exclusion chromatography, which pointed out that maltodextrins had a bimodal molecular weight distribution composed of high and low molecular weight oligo-saccharides. Indeed, maltodextrins are highly polydisperse materials (i.e. polydispersity index ranging from 5 to 12) and that should be the reason why such distinct behaviors were observed in some of the physico-chemical analyses that were preformed.