Polysaccharides, Volume 5, Issue 3 (September 2024) – 19 articles

Kefiran is a heteropolysaccharide that is considered a postbiotic and is obtained by kefir grains fermented in cow’s milk, while little is known about the donkey milk (DM) variety. Postbiotics are recognised as having important human health benefits that are very similar to probiotics but without the negative effects associated with their ingestion. Donkey is a monogastric animal, as are humans, and when used as an alternative food for infants who suffer from cow milk protein allergies, DM could therefore display more biocompatibility. In this study, the DM kefiran was extracted by ultrasound from kefir grains cultured in donkey milk and fully characterized for its structural and physicochemical properties by Fourier-transform infrared spectroscopy (FT-IR), High-Performance Liquid Chromatography- Refractive Index (HPLC-RI), Scanning electron microscope (SEM), Differential Scanning Calorimeters (DSC) and rheological analyses. In addition, tests were conducted on keratinocytes cell lines and human red blood cells to assess the nontoxicity and haemolysis degree of the polymer. The extraction yield of the DM kefiran was 6.5 ± 0.15%. The FT-IR analysis confirmed the structure of the polysaccharide by showing that the stretching of the C-O-C and C-O bonds in the ring, which formed two bands at 1157 and 1071 cm⁻¹, respectively, and the anomeric band at 896 cm⁻¹ indicates the β configuration and vibrational modes of glucose and galactose. Results were confirmed by HPLC-RI analysis indicating that the ratio glucose/galactose was 1:0.87. Furthermore, the SEM analysis showed a porous and homogeneous structure. The rheological analysis confirmed the pseudoplastic nature of the polymer, while the DSC analysis highlighted excellent thermal resistance (324 °C). Finally, DM kefiran was revealed to have biologically acceptable toxicity, showing a haemolytic activity of less than 2% when using fresh human red blood cells and showing no cytotoxicity on human keratinocytes. Therefore, kefiran obtained by DM shows an excellent biocompatibility, establishing it as a promising polymer for bioengineering human tissue for regenerative applications. Full article

Fenugreek (Trigonella foenum-graecum L.) is an annual, dicotyledonous medicinal plant which belongs to the Leguminosae family, and its leaves and seeds are widely used and cultivated throughout the world. Their widespread utilization is attributed to the great variety of primary and secondary metabolites they contain, such as flavonoids, alkaloids, steroidal saponins, tannins, as well as carbohydrates, in particular galactomannan, which is the focus of the current study. The presence of an equal number of galactose and mannose residues (Gal/Man ratio of 1:1) prevents the formation of hydrogen bonds between the mannose ones. This determines the good solubility of fenugreek galactomannan in cold water, even at low concentrations. The water solubility would be significantly better than that of carob and even slightly higher than that of guar gum, precisely due to their structural characteristics, which contribute to their possible advantages. Moreover, it is a good alternative as an excipient for the development of pharmaceutical dosage forms, as well as in the preparation of food products, affecting not only their structure but also their shelf life. Furthermore, it has promising applications not only in the fields of medicine and pharmaceutics but also offers environmental benefits. All of the above-mentioned factors are of high interest and qualify fenugreek galactomannan as a versatile polysaccharide, which is the reason for summarizing its benefits in this review. Full article

Cellulose obtained from oat hulls by bleaching with peracetic acid was modified, employing an ultrasound method that resulted in an esterification reaction with different vegetable oils (soybean, sunflower, and coconut) to produce modified cellulose (MC) with increased hydrophobicity. MC samples were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction, scanning electron microscopy, and their wettability and oil and water absorption capacities. FTIR indicated that the reaction occurred with all oils, which was observed by forming a new band associated with ester carbonyl groups at 1747 cm⁻¹. The modification did not affect the crystalline structure or surface morphology of the cellulose. MC samples modified with all oil sources showed a 6 to 9-fold decrease in water absorption capacity, a 3-fold increase in oil absorption capacity, and a higher affinity for nonpolar solvents. The modified samples adsorbed lower amounts of water at a slower rate. Different oil sources did not affect the main properties of MC. The ultrasonication-assisted process was not only effective in modifying cellulose by esterification with vegetable oils but was also an eco-friendly and simple strategy that does not require toxic reagents, providing reassurance of its sustainability. Full article

Protein hydrolysates containing bioactive peptides have emerged as therapeutic agents. However, these peptides may lose this bioactivity under gastrointestinal conditions. Encapsulation in edible biopolymers is a solution to this problem. Protein hydrolysates with ACE-I inhibitory activity, obtained previously, were encapsulated. A 1% solution of the biopolymers alginate (AG) and pectin (PC) in various ratios was prepared. The beads formed were evaluated in both wet and dry states for size, roundness, thermal gravimetric analysis (TGA), encapsulation efficiency, and ACE-I inhibitory activity. Selected samples underwent in vitro digestion, after which peptide release and ACE-I inhibitory activity were determined. Size analysis revealed that increasing the PC content increased the bead size, with 100% PC beads showing total deformation and reduced roundness. TGA indicated that wet beads had lower thermal stability compared to dry beads. The highest encapsulation efficiency (95.57% ± 0.49) was observed with 100% AG beads. The 75% AG 25% PC beads exhibited the highest ACE-I inhibitory activity (97.97% ± 1.01). Encapsulated protein hydrolysates retained their ACE-I inhibitory activity after simulated digestion, whereas non-encapsulated hydrolysates lost their bioactivity. Encapsulation of amaranth protein hydrolysates with AG and PC thus preserves antihypertensive activity even after in vitro digestion. Full article

Beta-glucan (β-glucan), a naturally occurring complex polysaccharide, has drawn attention for its diverse health benefits, including immune system modulation. β-glucan was extracted from two fungi, Monascus purpureus (Mp) and Monascus kaoliang (Mk), cultured in Saccharina japonica via submerged fermentation. The yield, solubility, total sugar, reducing sugar, protein content, Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Thermogravimetric analysis (TGA), Scanning Electron Microscopy (SEM), in vitro free radical scavenging activity, and cytotoxicity were analyzed. A significant yield of β-glucans, with the contents of 51.30 ± 1.54% in Mp and 44.24 ± 1.18% in Mk was observed on a dry weight basis. Water solubility slightly varied, measuring 36.25 ± 1.14% in Mp and 31.25 ± 0.94% in Mk. Total sugar and reducing sugar content in Mp and Mk derived β-glucans were 114.75 ± 2.54 mg/g and 100.25 ± 1.86 mg/g, 7.38 ± 0.78 mg/g, and 8.39 ± 0.46 mg/g, respectively. FTIR spectra resembled the standard, and TGA confirmed heat stability. XRD patterns indicated that the extracted β-glucans, including the standard one, showed the most prominent diffraction peaks in the lower 2θ range, suggesting similar crystalline phases; however, they differed in crystallinity and degree of amorphous content. SEM images displayed characteristic rough and fibrous shapes and surfaces for extracted β-glucans but it was uniform and of a regular shape in the standard sample. The isolated β-glucans exhibited in vitro free radical scavenging and no cytotoxicity was observed in the MTS assay. Therefore, utilizing S. japonica as a substrate in the fermentation process by Monascus spp. presents a unique opportunity in the production and utilization of β-glucans. Full article

The layer-by-layer (LbL) assembling of oppositely charged polyelectrolytes was studied using semi-synthetic polysaccharide derivatives, namely the polycations 6-aminoethylamino-6-deoxy cellulose (ADC) and cellulose (2-(ethylamino)ethylcarbamate (CAEC), as well as the polyanion cellulose sulfate (CS). The synthetic polymers poly(allylamine) (PAH) and poly(styrene sulfonate) (PSS) were employed as well for comparison. The stepwise adsorption process was monitored by whispering gallery mode (WGM) experiments and zeta-potential measurements. Distinct differences between synthetic- and polysaccharide-based assemblies were observed in terms of the quantitative adsorption of mass and adsorption kinetics. The LbL-approach was used to prepare µm-sized capsules with the aid of porous and non-porous silica particle templates. The polysaccharide-based capsule showed a switchable permeability that was not observed for the synthetic polymer materials. At ambient pH values of 7, low-molecular dyes could penetrate the capsule wall while no permeation occurred at elevated pH values of 8. Finally, the preparation of protein-loaded LbL-capsules was studied using the combination of CAEC and CS. It was shown that high amounts of protein (streptavidin and ovomucoid) can be encapsulated and that no leaking or disintegration of the cargo macromolecules occurred during the preparation step. Based on this work, potential use in biomedical areas can be concluded, such as the encapsulation of bioactive compounds (e.g., pharmaceutical compounds, antibodies) for drug delivery or sensing purposes. Full article

In recent decades, cellulose (Cel) and its modified forms have emerged as a new class of versatile adsorbents for removing dyes from aqueous solutions. This work reports the immobilization of macromolecules obtained from reactions between ethylenediamine (N) and ethylene sulfide (S) in three molar proportions (1:1, 1:2, and 1:4) on the surface of chlorinated cellulose (Cl-Cel), aiming to increase the adsorption capacity of dyes. The materials obtained (NS-Cel, N2S-Cel, and N4S-Cel) were characterized by elemental analysis, which demonstrated immobilization of macromolecules with a ratio of 12 ethylene sulfides to 1 ethylenediamine in the materials NS-Cel and N4S-Cel and a ratio of 10 ethylene sulfides to 1 ethylenediamine in NS-Cel. Intense C-H stretching bands of CH₂ groups at 2900 cm⁻¹ in the FT-IR spectra suggest a large amount of the functional group, corroborating the ¹³C NMR spectra, which presented a signal at 33 ppm referring to methylene carbons. The materials obtained had excellent performance in removing the dyes studied, with the adsorption capacity of the Remazol yellow GR dye being approximately 24 times greater than the raw material (87.70 ± 2.63 mg g⁻¹) for the best-hyperbranched cellulose N4S-Cel and 3.60 ± 0.18 mg g⁻¹ for Cel, and about ten times higher for the dye Remazol red RB (57.84 ± 1.73 mg g⁻¹) for N4S-Cel compared to previously published work for Cel. Full article

Gel electrophoresis and size exclusion chromatography (SEC) are vital techniques in biochemical research, employing gel matrix structures made of polysaccharides or synthetic polymers like polyacrylamide for the analysis and separation of macromolecules. Polysaccharides, such as agarose, offer safer alternatives to acrylamide. Polysaccharide gels, notably agarose, facilitate the analysis and purification of proteins and nucleic acids through a molecular sieving mechanism. Gel electrophoresis for proteins is mainly divided into denaturing and native methods. Denaturing electrophoresis with sodium dodecyl sulfate (SDS) simplifies protein migration but disrupts molecular interactions. Conversely, native gel electrophoresis, without SDS, allows proteins to migrate based on the running pH and the isoelectric point of the proteins, while nucleic acids consistently migrate toward the anode. The electrophoresis of proteins with variable charges presents complexes. This review focuses on the use of polysaccharides, particularly agarose, for native gel electrophoresis, highlighting their applications in separating macromolecules. It also discusses the applications and limitations of agarose gels when used as a matrix for electrophoresis. Such information should help in designing electrophoresis experiments using polysaccharides. Full article

Chitosan was subjected to a crosslinking reaction with three polyhydroxylated diacids (glucaric (GlcA), mannaric (ManA), and mucic (MucA) acids) that only differ in the spatial orientation of their hydroxyl groups. This work aimed to obtain experimental evidence of the impact of the three-dimensional arrangement of the crosslinkers on the resulting properties of the products. In all the cases, the products were hydrogels, and their chemical structures were fully elucidated by FT-IR spectroscopy and conductometric titration. Thermogravimetric and morphological studies were also carried out. The specific surface area of all the products was similar and higher than that of native chitosan. Moreover, all hydrogels were characterized in terms of viscoelastic properties and long-term stability under external perturbation. Furthermore, their lead adsorption efficiency and swelling capacity were assessed. Despite the resemblant chemical structure in all the hydrogels, Ch/ManA exhibited the highest lead adsorption capacity, (Ch/ManA: 93.8 mg g⁻¹, Ch/GlcA: 82.9 mg g⁻¹, Ch/MucA: 79.2 mg g⁻¹), while Ch/GlcA exhibited a remarkably higher swelling capacity (i.e., ~30% more than Ch/MucA and ~40% more than Ch/ManA). The results obtained herein evidenced that the selection of the polyhydroxylated crosslinker with the appropriate three-dimensional structure could be crucial to finely adjust the final materials’ features. Full article

Carboxymethylglucans (CMGs) are widely used semisynthetic polyelectrolytes, e.g., for pharmaceuticals. They are produced in heterogeneous processes on activated starch granules or cellulose fibers. In contrast to neutral ether derivatives, a lower DS in the range, commonly between 0.6 and 1.2, is sufficient to achieve the water solubility of CM cellulose. The high proportion of unsubstituted domains, which could aggregate and therefore only swell and form gel particles but do not dissolve, places higher demands on the statistical distribution of the substituents. The knowledge of regioselectivity, essential for the interpretation of higher structural-level data, can be obtained by various methods, preferentially by CE/UV after hydrolysis. To study the distribution of substituents at the polymer level by mass spectrometric (MS) analysis, partial random depolymerization is required. Due to the ionic character and acid functionality, all the attempts of the direct depolymerization of CMG and further sample preparation suffered from bias, side reactions, and multiple ion formation in MS. Finally, the transformation of CMGs to the corresponding hydroxyethylglucans (HEGs) by the reduction of the esterified carboxy groups with LiAlH₄ opened the window for quantitative oligomer MS analysis. While the CM amyloses were reduced quantitatively, the transformation of the CMC was only about 60% but without the formation of mixed CM/HE ethers. Full article

Tunable hydrogels have gained significant attention in the bioengineering field due to their designer preparation approach. Towards this end, gelatine stands out as a promising candidate owing to its desirable attributes, such as biocompatibility, ability to support cell adhesion and proliferation, biodegradability, and cost-effectiveness. This study presents the preparation of a robust gelatine hydrogel employing sacran aldehyde (SDA) as a natural cross-linker. The resulting SDA-cross-linked gelatine hydrogels (GSDA) display an optimal compressive stress of 0.15 MPa at 50% strain, five times higher than pure gelatine hydrogel. As SDA cross-linking concentration is increased, the swelling capacity of GSDA declines. This decline in swelling capacity, from 80% to 40%, is a result of strong crosslinking of gelatin with SDA. Probing further with FT-IR spectroscopy and SEM at the micron scale unveiled a dual-cross-linking mechanism within the hydrogels. This mechanism encompasses both short- and long-range covalent cross-linking, along with thermo-induced physical cross-linking, resulting in a significant enhancement of the load-bearing capacity of the fabricated hydrogels. Full article

This review explores the production of the fungal polysaccharide pullulan by mutants and natural isolates of Aureobasidium species using strain improvement. Pullulan is a neutral polysaccharide gum whose structure is a maltotriose-containing glucan. This polysaccharide gum has applications in the fields of food, pharmaceuticals, biomedical and wastewater treatment. The strain improvement of Aureobasidium species has focused on the pullulan production process, including the isolation of strains exhibiting reduced pigmentation, polysaccharide overproduction, the production of pullulan with variable molecular weight, and increased osmotolerant strains promoting pullulan production at high carbon source concentrations and pullulan production on hemicellulosic substrates. The majority of studies have emphasized the isolation of reduced pigmentation and pullulan hyperproducer strains since the goal of large-scale commercial pullulan production is to synthesize non-pigmented polysaccharides. A promising area of strain improvement is the isolation of strains that synthesize authentic pullulan from hemicellulosic substrates. If strain improvement in this area is successful, the goal of commercially producing pullulan at a competitive cost will eventually be achieved. Full article

This study focused on utilizing cellulose nanocrystal (CNC)–polyvinyl alcohol (PVA) composite in optical sensor applications to detect high humidity conditions and determine water concentration in ethanol. We focused on the composite’s effectiveness in moisture absorption to demonstrate visual color change. We demonstrated that the different molecular weights of PVA significantly affect CNC’s chiral nematic structure and moisture absorption capability. PVA with molecular weight 88 k–97 k exhibited the disintegration of its chiral nematic structure at 30 wt%, whereas low molecular weight PVA (n~1750) showed no structural disintegration even at 100 wt% concentration when analyzed through UV-Vis spectroscopy. Further, the thermal crosslinking of the CNC-PVA composite showed no significant loss of moisture sensitivity for all molecular weights of the PVA. We observed that the addition of PVA to the sulfated CNC obtained from sulfuric acid hydrolysis did not facilitate moisture absorption significantly. A CNC-PVA sensor was developed which can detect high humidity with 2 h. of exposure time. 2,2,6,6-tetramethylpiperidin-1-piperidinyloxy oxidized CNC (TEMPO-CNC) having carboxylic functionality was also used to prepare the CNC-PVA composite films for comparing the effect of functional groups on moisture sensitivity. Finally, we demonstrated a facile method for utilizing the composite as an optical sensor to detect water concentration in ethanol efficiently; thus, it can be used in polar organic solvent dehydration applications. Full article

Exopolysaccharides (EPSs) are a diverse class of biopolymers synthesized by microorganisms under environmental stress conditions, such as pH, temperature, light intensity, and salinity. They offer biodegradable and environmentally friendly alternatives to synthetic polymers. Their structural versatility and functional properties make them unique in various industries, including food, pharmaceuticals, biomedicine, cosmetics, textiles, petroleum, and environmental remediation. In this way, among the well-known EPSs, homopolysaccharides like dextran, bacterial cellulose, curdlan, and levan, as well as heteropolysaccharides like xanthan gum, alginate, gellan, and kefiran, have found widespread applications in numerous fields. However, recent attention has focused on the potential role of extremophile bacteria in producing EPSs with novel and unusual protective and biological features under extreme conditions. Therefore, this review provides an overview of the functional properties and applications of the commonly employed EPSs. It emphasizes their importance in various industries and scientific endeavors while highlighting the raised interest in exploring EPSs with novel compositions, structures, and properties, including underexplored protective functionalities. Nevertheless, despite the potential benefits of EPSs, challenges persist. Hence, this review discusses these challenges, explores opportunities, and outlines future directions, focusing on their impact on developing innovative, sustainable, and functional materials. Full article

Glycogen is a natural polysaccharide used as an energy storage macromolecule. The role of glycogen metabolism in type 2 diabetes mellitus has been under investigation for several years, along with its implication in cancer and cardiovascular and neurodegenerative diseases. Previous studies using pig liver glycogen with rabbit muscle glycogen phosphorylase (RMGPb), which catalyzes the first step of glycogen degradation to glucose-1-phosphate, showed that the surface of an average glycogen molecule is covered by a total of 20 RMGPb dimeric molecules. In this work, we selected oyster glycogen (Glyc) to investigate its interaction with RMGPb by employing biophysical techniques. Dynamic, static, and electrophoretic light scattering were used to investigate the solution behaviors and structures of both the Glyc molecule itself and the formed complexes between Glyc and GPb at different mixing ratios. It was established that the interaction between oyster Glyc and RMGPb is similar to that previously reported for pig liver glycogen. Moreover, the structure of the complexed GPb was monitored by fluorescence and FTIR spectroscopy. Full article

Mushroom polysaccharides, key components of fungal cell walls, exhibit various biological properties and hold significant medicinal and industrial value. These polysaccharides are known for their medicinal properties like antitumor, antioxidant, anticancer, immunomodulatory, and antiviral properties. Mushroom polysaccharides, particularly β-glucans, α-glucans, and chitin, have been associated with various health benefits. β-glucans are well studied for their bioactivities, while α-glucans and chitin have gained attention for their prebiotic, antimicrobial, and wound-healing properties. The therapeutic effects of these polysaccharides are closely linked to their chemical structures, including molecular weight, monosaccharide composition, and glycosidic bond types. This work aims to review the studies on mushroom polysaccharides, with a particular focus on their structural composition to deepen medicinal properties of mushroom polysaccharides. Also, the extraction methods and the pharmaceutical application of polysaccharides will be revised in this work. Full article

Carboxymethyl cellulose sodium salt is a common water-soluble derivative of cellulose. It serves as a bioinert mucoadhesive material extensively used in biomedicine, particularly for crafting targeted drug delivery systems. In our study, we demonstrate that graft copolymers of sodium carboxymethyl-cellulose with poly(N-vinylimidazole) can function as carriers for the antibacterial drug metronidazole. Non-covalent associations form between the components, excluding the involvement of the nitro groups of the drug in the interaction. These loaded copolymers exhibit the capability to release the drug under conditions mimicking the stomach environment for up to 48 h. This renders the obtained associations promising candidates for the development of a metronidazole-targeted delivery system. Full article

The goal of this study was to explore a route for introducing functionalities into agarose-based hydrogels to tune the physical, chemical, and biological properties. Several agarose derivatives were prepared by homogeneous synthesis, including anionic agarose sulfates (ASs), reactive azido agaroses (AZAs), and cationic agarose carbamates (ACs), as well as agarose tosylates (ATOSs) and agarose phenyl carbonates (APhCs). The products were characterized in terms of their molecular structure and solubility behavior. The results suggest that the native gel-forming ability of agarose is retained if the introduced functionalities are hydrophilic, and the overall degree of substitution is low (DS < 0.5). Thus, functional hydrogels from several agarose derivatives could be obtained. The mechanical stability of the functional hydrogels was decreased compared to native agarose gels but was still in a range that enables safe handling. An increase in mechanical strength could be achieved by blending functional agarose derivatives and agarose into composite hydrogels. Finally, it was demonstrated that the novel functional agarose hydrogels are biocompatible and can potentially stimulate interactions with cells and tissue. Full article

Natural fibers have been widely employed in different fields as composite materials. However, the thermal behavior of natural materials is not fully understood since the chemical components chemically interact with each other. Put simply, the thermal degradation kinetics of natural fibers are of crucial importance in the academic and industrial fields. This study objectively fully investigated the thermal behavior of different natural fibers using the thermal kinetic method criteria, as described by the ICTAC (International Committee of Thermal Analysis and Calorimetry). The kinetic triplet (activation energy, pre-exponential factor, and reaction mechanism) was determined as a function of the conversion degree. For all plant fibers, the results indicated an autocatalytic process with an average activation energy and pre-exponential factor of 200 kJ·mol⁻¹ and 40 s⁻¹, respectively. The activation energy can be mainly attributed to the cellulose component, while the pre-exponential factor is due to the possible compensation effect as a mathematical artifact of the Arrhenius-based equation. Finally, the lifetime prediction of the plant fibers was estimated using the pre-determined kinetic triplet parameters to estimate the plant fiber stability under isothermal conditions. Full article