Alcohol Precipitation Research Articles

Rumex japonicus Houtt. Leaves: the chemical composition and anti-fungal activity

BackgroundCandida albicans is a pathogenic commensal fungus. Trichophyton mentagrophytes and Trichophyton rubrum are the leading pathogens of dermatophysis. Rumex japonicus Houtt. has a miraculous effect on the treatment of tinea skin disease, but its mechanism has not been clarified. Purpose: This paper investigated the anti-fungal ingredients of the leaves of Rumex japonicus Houtt. (RJH-L) and the mechanism of the anti-fungal (Trichophyton mentagrophytes, Trichophyton rubrum and Candida albicans). MethodFirst, the chemical composition analysis of RJH-L was conducted by acid extraction and alcohol precipitation, high performance liquid chromatography (HPLC) and nuclear magnetic resonance spectroscopy (NMR); in vitro anti-fungal experiments were carried out, including the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) spore germination rate, germ tube production rate, nucleic acid and protein leakage rate, biofilm structure, PCR, etc., to study the mechanism of action of RJH-L anti-fungal and anti-biofilm activity. ResultSeven monomer compounds were obtained: anthraquinones (rhein, emodin and aloe-emodin); polyphenols (ferulic acid, p-coumaric acid), and flavonoids (rutin and quercetin). The results of in vitro anti-fungal experiments showed that the extracts of RJH-L had strong inhibitory effect on both fungi (MIC: 1.96 µg/mL-62.50 µg/mL), of which emodin had the strongest effect on Trichophyton mentagrophytes; and rhein had the strongest effect on Candida albicans and Trichophyton rubrum. The above active components can inhibit the germination of fungal spores and germ tube, change cell membrane permeability, prevent hyphal growth, destroy the biofilm structure, and down-regulate the expression of agglutinin-like sequencefamily1 of biofilm growth. ConclusionThis study shows that RJH-L are rich in polyphenols, flavonoids, and anthraquinones, and play a fungicidal role.

In vitro evaluation of the antitumor and antioxidant effects of purified and characterized polysaccharides from Ganoderma applanatum.

In this study, the chemical properties of Ganoderma applanatum polysaccharides (GAP) were systematically investigated, followed by a comprehensive analysis of their antitumor and antioxidant capabilities. Ultrasonic circulation technology was employed for the extraction of GAP, facilitating the procurement of crude polysaccharides through the Sevag method, dialysis, and sequential alcohol precipitation. The chemical constituents of these polysaccharides were subsequently analyzed utilizing Fourier-transform infrared spectroscopy and gas chromatography-mass spectrometry. The antitumor and antioxidant properties of the polysaccharide components were assessed utilizing 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and free radical scavenging methods, respectively. Gradient ethanol precipitation yielded three principal polysaccharide fractions: GAP-40, GAP-60 and GAP-80. Analysis revealed glucose as the predominant component in these fractions, with average molecular weights determined to be 77.75, 9.25 and 1.03 kDa, respectively. The antitumor activity of GAP-40, GAP-60 and GAP-80 against MCF-7 cells demonstrated both time and concentration dependence. Significantly, GAP-40, at a concentration of 1000 μg/mL over 48 h, presented a notable inhibition rate of 56.77%, outperforming GAP-60 and GAP-80. Furthermore, the antioxidant capacities of GAP-40, GAP-60 and GAP-80 were comparably significant to that of vitamin C, with detailed analysis revealing marked differences in antioxidant activity among the GAP variants. Specifically, GAP-40 exhibited superior efficacy in scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) radicals relative to GAP-60 and GAP-80. In contrast, GAP-80 was distinguished by its exceptional hydroxyl radical scavenging capacity, surpassing that of both GAP-40 and GAP-60. These results substantiate the potential of GAPs as viable and effective antitumor agents, additionally suggesting their utility as functional foods endowed with antioxidant attributes.

Optimization of extraction process of polysaccharide from Phylloporia fontanesiae and its simulated digestion in vitro.

In this study, Phylloporia fontanesiae polysaccharide was successfully isolated through a sequential water extraction and alcohol precipitation process. Utilizing the Box-Behnken design, the extraction process was optimized based on single-factor experiments, considering variables such as the material-to-liquid ratio, extraction temperature, extraction time, and the number of extractions. The polysaccharide composition of P. fontanesiae is predominantly composed of mannose, glucuronic acid, glucose, and galactose, with a molar mass ratio of 4.31:4.10:36.83:1, along with minor amounts of aminoglucose and fucose. The polysaccharide fraction of P. fontanesiae comprises two distinct components, possessing relative molecular masses of 8.85kDa and 134.03kDa. Notably, the polysaccharide exhibited significant antioxidant activity. After undergoing simulated gastrointestinal digestion, no significant changes were observed in its antioxidant activity, molecular weight, or monosaccharide composition. This study not only enhanced the extraction efficiency of P. fontanesiae polysaccharide but also provided valuable insights into its composition, structure, and digestion characteristics. PRACTICAL APPLICATION: The optimum extraction process, stability, and antioxidant activity of Phylloporia fontanesiae polysaccharide during simulated digestion of gastrointestinal tract were studied. The results provide a theoretical basis for the development and application of this polysaccharide in the field of food and health products.

The structural features and anti-inflammatory properties of a glucogalactan from Holotrichia diomphalia Bates (Qi Cao)

Ethnopharmacological relevanceThe dried larvae of Holotrichia diomphalia Bates, named Qi Cao, is a traditional Chinese medicine treat for liver diseases and arthritis. Polysaccharides is a principal component in Qi Cao, which exhibiting antioxidant and anti-inflammatory effects. However, the structural characteristics and underlying mechanisms of the polysaccharides remain inadequately elucidated. Aim of the studyTo analyze the primary structure and elucidate the molecular anti-inflammatory mechanisms of the active polysaccharide in Qi Cao. Materials and methodsThe total polysaccharide was extracted by water extraction and alcohol precipitation, and further isolated and purified by DEAE Sephadex A-25 column and Sephadex G-100 column. The anti-inflammatory properties of four major fractions (HDPS-1, HDPS-2, HDPS-3, HDPS-4) and the pure homogeneous polysaccharides (HDPS-1I and HDPS-1II) were assessed using a RAW 264.7 cell model induced by lipopolysaccharide (LPS), and HDPS-1II was identified as the polysaccharide exhibiting significant anti-inflammatory activity in Qi Cao. The structural characteristics of HDPS-1II were subsequently analyzed using high-performance size-exclusion chromatography (HPSEC), fourier-transform infrared spectroscopy (FT-IR), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) spectroscopy. The TLR4, NF-κB, COX-2 and iNOS expressions were determined by Western blot analysis to investigate the anti-inflammatory mechanism of HDPS-1II in vitro. Finally, the in vivo anti-inflammatory activity of HDPS-1II were evaluated by measuring the serum levels of pro-inflammatory factors, inflammatory cell infiltration and organelle damage in the lung tissues of sepsis model mice. ResultsA homogeneous polysaccharide (HDPS-1II) with molecular weight of 1.7 × 104 Da was isolated from Holotrichia diomphalia Bates. HDPS-1II contains a backbone of α-T-Glcp-(1 → 6)-α-Glcp-(1 → 4)-α-Galp-(1 → 4)-α-Galp-(1 → 6)-α-Galp-(1 → 3)-α-Galp-(1 → . It inhibited activation of the TLR4/NF-κB signaling and reduced pro-inflammatory factors and NO in LPS-stimulated macrophage. Moreover, HDPS-1II increased the survival rate, inhibited inflammatory cells infiltration, and ameliorated the lung tissue damage in septic mice. ConclusionsHDPS-1II exhibits anti-inflammatory effects in vitro and in vivo, which is the active polysaccharide components of the anti-inflammatory activity of Qi Cao.

Hypolipidemic Effects of Mesona chinensis Benth Polysaccharides with Different Structures and Molecular Weights.

Four novel Mesona chinensis Benth polysaccharides were isolated using aqueous alcohol precipitation. Their molecular weights were determined using high-performance gel permeation chromatography: MA1 (2.3 kDa), MA2 (80.5 kDa), MA3 (180.9 kDa), and MA4 (635.2 kDa), and their compositions were analyzed using GC-MS. The polysaccharides were mainly D-glucose, D-galactose, L-Rhamnose, D-arabinose, D-xylose, and D-mannose. The structural characteristics were further analyzed using infrared spectrophotometry and were identified as a type of pyrrhic sugar. An insulin-induced insulin resistance model of HepG2 cells and oleic acid-induced fat accumulation model of insulin were established to evaluate the hypolipidemic effects. Three Bacteroides spp. [Bacteroides thetaiotaomicron (BT), B. ovatus (BO), and B. cellulosilyticus (BC)] that were negatively correlated with lipid-lowering activity were used to evaluate the lipid-lowering activity of polysaccharides. The Bacteroides metabolites of MA1 and MA2 exhibited hypolipidemic effects and antioxidant activities and could potentially be used as lipid-lowering supplements.

Tailoring highland barley bran arabinoxylans: Insights into composition, structure, and functional properties

Highland barley bran (HBB) stands as an underutilized source of arabinoxylans (AXs), presenting promising prospects for various applications. This study focuses on the isolation of four different AXs (S20, S40, S60, S80) from HBB using alkali extraction and gradient alcohol precipitation. These AXs exhibited diverse purity (56.47%–80.92%), monosaccharide compositions (A/X:0.47–0.96), and molecular weights (Mw) (173.5–498.2 kDa) of different AXs, along with different microstructural variations, amorphous qualities, and thermal stability. Rheological analyses demonstrated their shear-thinning behavior and distinctive gel-forming capacities, with S20 exhibiting superior viscosity and gel-forming ability. Notably, all four AXs demonstrate notable DPPH and ABTS radical scavenging capacities, with S80 demonstrating the highest antioxidant potency. Correlation analyses highlighted connections between molecular weight, branching degree, and rheological and antioxidant properties of AXs. Among different AXs, S20, with its low branching degree (0.47) and high molecular weight (390.4 kDa), exhibited significant viscosity, suggesting its suitability as a natural thickener in food processing. Conversely, S80, characterized by its low molecular weight (173.5 kDa), high branching degree (0.96), and potent antioxidant activity, holds promise as a natural antioxidant in functional foods. This study provides crucial insights for tailoring AX applications, unlocking the potential for the valorization of HBB in diverse scenarios.

Structural characteristics and anti-tumor activity of alcohol-precipitated polysaccharides from Cordyceps militaris under different ethanol concentrations

Six Cordyceps militaris polysaccharides (named CMP-1, CMP-2, CMP-3, CMP-4, CMP-9, and CMP-A) were obtained by fractional alcohol precipitation. The experimental results showed that the six Cordyceps militaris polysaccharides had similar chemical composition and spectral features, and different molecular weights, monosaccharide compositions and anti-tumor activities. Purification of CMP-9 yielded the small molecule polysaccharide LMW-CMP (3.06 kDa). Structural experiments showed that LMW-CMP is an α-glucan with (1 → 4)-α-D-Glcp as the main chain and a glucose branched chain attached at the O-6 position. The results of cell experiments showed that LMW-CMP could effectively inhibit the growth and proliferation of HepG2 cells, activate the downstream NF-κB signaling pathway through the MAPK pathway to induce apoptosis of HepG2 cells, and block apoptosis at the G1 phase. Animal experiments showed that LMW-CMP inhibited the proliferation of tumor cells in H22 tumor-bearing mice by improving the state of immune organs, increasing the activity of immune cells and cytokine levels in the body, and regulating the distribution of lymphocyte subpopulations, with a tumor inhibition rate of 45.70 % (200 mg/kg).

Novel manufacturing process of pneumococcal capsular polysaccharides using advanced sterilization methods.

Pneumococcal disease is caused by Streptococcus pneumoniae, including pneumonia, meningitis and sepsis. Capsular polysaccharides (CPSs) have been shown as effective antigens to stimulate protective immunity against pneumococcal disease. A major step in the production of pneumococcal vaccines is to prepare CPSs that meet strict quality standards in immunogenicity and safety. The major impurities come from bacterial proteins, nucleic acids and cell wall polysaccharides. Traditionally, the impurity level of refined CPSs is reduced by optimization of purification process. In this study, we investigated new aeration strategy and advanced sterilization methods by formaldehyde or β-propiolactone (BPL) to increase the amount of soluble polysaccharide in fermentation supernatant and to prevent bacterial lysis during inactivation. Furthermore, we developed a simplified process for the CPS purification, which involves ultrafiltration and diafiltration, followed by acid and alcohol precipitation, and finally diafiltration and lyophilization to obtain pure polysaccharide. The CPSs prepared from formaldehyde and BPL sterilization contained significantly lower level of residual impurities compared to the refined CPSs obtained from traditional deoxycholate sterilization. Finally, we showed that this novel approach of CPS preparation can be scaled up for polysaccharide vaccine production.

Extraction and Characterization of Pectin from Orange Peel

Fruit and vegetable processing waste contributes to around 30% of the total weight of the fruits and vegetables. These wastes can be converted into valuable goods for potential benefits. Therefore, this study was focused on extracting pectin from agricultural waste (orange peels) using two methods: alcohol precipitation and enzymatic extraction. The extracted pectin was compared in terms of various properties. Alcohol precipitation resulted in higher yield percentages (16.99 ± 0.8 % wet; 6.46 ± 1.01 % dry respectively) compared to the enzymatic method (16.84 ± 0.42 % wet; 3.66 ± 0.34 % dry, respectively). FT-IR and XRD spectra indicated the presence of highly methylated semi-crystalline pectin in both methods with potent antioxidant property (40.57%). This research work provides comparative extraction method of pectin from agricultural waste i.e. orange peels which has diverse applications in the food, pharmaceutical industries and biomedical applications.

Study on the chemical composition and anti-fungi activities of anthraquinones and its glycosides from Rumex japonicus Houtt.

Fungi, such as Trichophyton rubrum (T. rubrum) and Microsporum canis Bodin Anamorph (M. canis Bodin Anamorph) are the main pathogens of dermatophysis. According to ancient books records, Rumex japonicus Houtt. (RJH) has a miraculous effect on the treatment of dermatophysis. To reveal the anti-fungi (T. rubrum and M. canis Bodin Anamorph) components and its mechanism of the Rumex japonicus Houtt. The vinegar extraction and alcohol precipitation, HPLC and nuclear magnetic resonance spectroscopy (NMR) were employed for analyzing the chemical compositions of RJH; in vitro anti-fungal experiment was investigated including test the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC), spore germination rate, nucleic acid, protein leakage rate, biofilm structure, and the mechanism of anti-fungal and anti-fungal biofilms in RJH. Seven anthraquinones and their glycoside compounds were obtained in this study respectively, such as chrysophanol, physcion, aloe-emodin, emodin, rhein, emodin-8-O-β-D-glucoside and chrysophanol-8-O-β-D-glucoside. In vitro anti-fungal experiment results showed that RJH extracts have good anti-fungal activity for dermatophytic fungi. Among them, the MIC of the rhein, emodin and aloe-emodin against T. rubrum are 1.9µg/ml, 3.9µg/ml and 15.6µg/ml, respectively; the MIC of emodin and aloe-emodin against M. canis Bodin Anamorph are 7.8µg/ml and 62.5µg/ml, respectively. In addition, its active components can inhibit fungal spore germination and the formation of bud tube, change cell membrane permeability, prevent hyphal growth, destroy biofilm structure, and down-regulate the expression of agglutinin-like sequence family 1 of the adhesion phase of biofilm growth. The study shows that RJH play a fungicidal role.

Immunomodulatory Effect of Cordyceps militaris Polysaccharide on RAW 264.7 Macrophages by Regulating MAPK Signaling Pathways.

Polysaccharide is one of the principal bioactive components found in medicinal mushrooms and has been proven to enhance host immunity. However, the possible mechanism of immunomodulatory activity of Cordyceps militaris polysaccharide is not fully understood. Hot water extraction and alcohol precipitation, DEAE-Sephadex A-25 chromatography, and Sephadex G-100 chromatography were used to isolate polysaccharide from C. militaris. A high-molecular-weight polysaccharide isolated from C. militaris was designated as HCMP, which had an Mw of 6.18 × 105 Da and was composed of arabinose, galactose, glucose, mannose, and xylose in a mole ratio of 2.00:8.01:72.54:15.98:1.02. The polysaccharide content of HCMP was 91.2% ± 0.16. The test in vitro showed that HCMP activated mouse macrophage RAW 264.7 cells by enhancing phagocytosis and NO production, and by regulating mRNA expressions of inflammation-related molecules in RAW 264.7 cells. Western blotting revealed that HCMP induced the phosphorylation of mitogen-activated protein kinases (MAPKs). Moreover, using inhibitors of MAPKs decreased the mRNA levels of inflammation-related molecules induced by HCMP. These data evidenced that the immunomodulatory effect of HCMP on RAW 264.7 macrophages was mediated via the MAPK signaling pathway. These findings suggested that HCMP could be developed as a potent immunomodulatory agent for use in functional foods and dietary supplements.

Polysaccharides as Quality Marker to Rapid Profile for Ophiocordyceps sinensis by PXRD.

Ophiocordyceps sinensis has long been recognized as a mysterious and valuable traditional Chinese medicine but there has been little research on quality markers for O. sinensis. This study looked into the potential of using powder X-ray diffractometry (PXRD) to analyze polysaccharides as a quality marker for O. sinensis. There were 16 different habitats of O. sinensis collected in Qinghai, Gansu, Sichuan, Yunnan, and Tibet. In addition, five different types of Cordyceps species were collected. The characteristic diffraction peaks of O. sinensis were determined and then matched with the characteristic diffraction peaks of intracellular polysaccharides obtained from O. sinensis to determine the attribution relationship of the characteristic diffraction peaks. O. sinensis powder's X-ray diffraction pattern is determined by its composition, microcrystalline crystal structure, intramolecular bonding mechanism, and molecular configuration. After fractionation and alcohol precipitation of crude intracellular polysaccharide, mycelium crude intracellular polysaccharide (MCP) and fruiting body crude intracellular polysaccharide (FCP) were obtained and the fingerprint of O. sinensis was identified by the specific characteristic peaks of the X-ray diffraction pattern from intracellular polysaccharide. The results indicated that the PXRD patterns of different populations of O. sinensis were overlaid well with 18 characteristic diffraction peaks obtained by microcrystalline diffraction. Moreover, the powder diffractograms as a fingerprint provided a practical identification of O. sinensis from other Cordyceps species. In addition, we detected that the powder diffractograms of intracellular polysaccharide MCP and MCP75 could be coupled with the PXRD of O. sinensis. Specifically, 18 characteristic diffraction peaks were identified as coming from MCP and MCP75 according to those interplanar crystal spacing, which matched well with those of PXRD of O. sinensis. PXRD spectra combined with an updated multivariable discriminant model were found to be an efficient and sensitive method for O. sinensis quality control. According to the findings of this study, PXRD should be further investigated for quality control assessments and plant extract selection trials.

Structural characterization of three acidic polysaccharides from Opuntia dillenii Haw. fruits and their protective effect against hydrogen peroxide-induced oxidative stress in Huh-7 cells

Three novel acidic polysaccharide fractions (OFPP-1, OFPP-2, OFPP-3) with different molecular weights (803.7, 555.1 and 414.5 kDa) were isolated from the peeled Opuntia dillenii Haw. fruits by alkali-extraction, graded alcohol precipitation and column chromatography. Structural analysis indicated that OFPPs were pectic polysaccharides consisting of rhamnose, arabinose and galactose residues. The backbone of OFPP-1 consisted of a repeating unit →6-α-D-GalpA-(1→2)-α-L-Rhap-(1→ with T-α-D-GalpA-(1→6)-α-D-GalpA-(1→4)-α-D-Glcp-(1→, T-β-D-Xylp-(1→6)-α-D-GalpA-(1→4)-α-D-Glcp-(1→ or T-α-D-GalpA-(1→3)-α-L-Araf-(1→ as the side chains. The backbone of OFPP-2 consisted of a disaccharide repeating unit →2)-α-L-Rhap-(1→4)-β-D-GalpA-(1→ with T-β-L-Araf-(1→ as the branches substituted at the O-4 position of →2,4)-α-L-Rhap-(1→. Whereas the backbone of OFPP-3 was →2,4)-α-L-Rhap-(1→2)-α-L-Rhap-(1→3)-β-L-Araf-(1→ or →2,4)-α-L-Rhap-(1→2)-α-L-Rhap-(1→4)-β-D-GalpA-(1→, which was branched at the O-4 position of →2,4)-α-L-Rhap-(1→. Moreover, these 3 polysaccharide fractions could protect Huh-7 cells against H2O2-induced oxidative stress to different extents by decreasing the MDA content and increasing the SOD, CAT, GSH-Px activities and the GSH level in the Huh-7 cells. These results suggest that OFPPs have the potential to be used as natural antioxidants.

Steam Explosion Pretreatment of Polysaccharide from Hypsizygus marmoreus: Structure and Antioxidant Activity.

This paper investigated the effects of steam explosion (SE) pretreatment on the structural characteristics and antioxidant activity of Hypsizygus marmoreus polysaccharides (HPS). Hypsizygus marmoreus samples were pretreated at different SE temperatures (120-200 °C) and polysaccharides were extracted using the water extraction and alcohol precipitation method. The results showed that SE pretreatment improved the extraction rate of HPS. Under the conditions of SE treatment time of 60 s and temperature of 160 °C, the extraction rate of HPS was the highest (8.78 ± 0.24%). After SE pretreatment, the structural changes of HPS tended to enhance the antioxidant activity, which showed that the content of Gal and Man in the monosaccharide composition increased and the molecular weight decreased. When testing antioxidant activity in vitro, the ability of SE-pretreated HPS to scavenge DPPH radicals, hydroxyl radicals, and superoxide anion radicals was better than that of HPS without SE pretreatment. Our findings shed light on SE pretreatment as an efficient method for extracting active polysaccharides, providing a new way to improve their extraction rate and biological activity.

Recent Advances in Polysaccharides from Chaenomeles speciosa (Sweet) Nakai.: Extraction, Purification, Structural Characteristics, Health Benefits, and Applications.

This article systematically reviews the extraction and purification methods, structural characteristics, structure-activity relationship, and health benefits of C. speciosa polysaccharides, and their potential application in food, medicine, functional products, and feed, in order to provide a useful reference for future research. Chaenomeles speciosa (Sweet) Nakai. has attracted the attention of health consumers and medical researchers as a traditional Chinese medicine with edible, medicinal, and nutritional benefits. According to this study, C. speciosa polysaccharides have significant health benefits, such as anti-diaetic, anti-inflammatory and analgesic, anti-tumor, and immunomodulatory effects. Researchers determined the molecular weight, structural characteristics, and monosaccharide composition and ratio of C. speciosa polysaccharides by water extraction and alcohol precipitation. This study will lay a solid foundation for further optimization of the extraction process of C. speciosa polysaccharides and the development of their products. As an active ingredient with high value, C. speciosa polysaccharides are worthy of further study and full development. C. speciosa polysaccharides should be further explored in the future, to innovate their extraction methods, enrich their types and biological activities, and lay a solid foundation for further research and development of products containing polysaccharides that are beneficial to the human body.

Structural characterization and innate immunomodulatory effect of glucomannan from Bletilla striata

The crude polysaccharide of Bletilla striata in this study was extracted by water extraction and alcohol precipitation and further purified by gel column to yield the purified component Bletilla striata polysaccharide (BSP). Its structure and innate immune regulation activity were studied. BSP mainly comprises mannose and glucose, with a monosaccharide molar ratio of 2.9:1 and a weight-average molecular weight of 28,365 Da. It is a new low-molecular-weight water-soluble neutral glucomannan. BSP contains a → 6)-β-Manp-(1→, →4)-β-Glcp-(1→, →4)-β-Manp-(1 → and →3)-α-Manp-(1 → linear main chain, containing β-Glcp-(1 → and β-Manp-(1 → two branched chain fragments were connected to the Man residue at position 4. BSP can enhance the anti-infection ability of Caenorhabditis elegans against Pseudomonas aeruginosa, significantly improve the phagocytic ability of RAW264.7 macrophages, stimulate the secretion of NO and TNF-α, and have good innate immune regulation activity. These findings guide the use of Bletilla striata polysaccharides with immunomodulatory action.

Glycomacropeptide: A comprehensive understanding of its major biological characteristics and purification methodologies.

Glycomacropeptide (GMP) is a bioactive peptide derived from whey protein, consisting of 64 amino acids. It is a phenylalanine-free peptide, making it a beneficial dietary option for individuals dealing with phenylketonuria (PKU). PKU is an inherited metabolic disorder characterized by high levels of phenylalanine in the bloodstream, resulting from a deficiency of phenylalanine dehydrogenase in affected individuals. Consequently, patients with PKU require lifelong adherence to a low-phenylalanine diet, wherein a significant portion of their protein intake is typically sourced from a phenylalanine-free amino acid formula. GMP has several nutritional values, numerous bioactivity properties, and therapeutic effects in various inflammatory disorders. Despite all these features, the purification of GMP is an imperative requirement; however, there are no unique methods for achieving this goal. Traditionally, several methods have been used for GMP purification, such as thermal or acid treatment, alcoholic precipitation, ultrafiltration (UF), gel filtration, and membrane separation techniques. However, these methods have poor specificity, and the presence of large amounts of impurities can interfere with the analysis of GMP. More efficient and highly specific GMP purification methods need to be developed. In this review, we have highlighted and summarized the current research progress on the major biological features and purification methodologies associated with GMP, as well as providing an extensive overview of the recent developments in using charged UF membranes for GMP purification and the influential factors.

Purple sweet potato polysaccharide ameliorates concanavalin A-induced hepatic injury by inhibiting inflammation and oxidative stress

BackgroundAutoimmune hepatitis (AIH) is a prevalent liver disease that can potentially lead to hepatic fibrosis and cirrhosis. The prolonged administration of immunosuppressive medications carries significant risks for patients. Purple sweet potato polysaccharide (PSPP), a macromolecule stored in root tubers, exhibits anti-inflammatory, antioxidant, immune-enhancing, and intestinal flora-regulating properties. Nevertheless, investigation into the role and potential mechanisms of PSPP in AIH remains notably scarce. PurposeOur aim was to explore the possible protective impacts of PSPP against concanavalin A (Con A)-induced liver injury in mice. MethodsPolysaccharide was isolated from purple sweet potato tubers using water extraction and alcohol precipitation, followed by purification through DEAE-52 cellulose column chromatography and Sephadex G-100 column chromatography. A highly purified component was obtained, and its monosaccharide composition was characterized by high performance liquid chromatography (HPLC). Mouse and cellular models induced by Con A were set up to investigate the impacts of PSPP on hepatic histopathology, apoptosis, as well as inflammation- and oxidative stress-related proteins in response to PSPP treatment. ResultsThe administration of PSPP significantly reduced hepatic pathological damage, suppressed elevation of ALT and AST levels, and attenuated hepatic apoptosis in Con A-exposed mice. PSPP was found to mitigate Con A-induced inflammation by suppressing the TLR4-P2X7R/NLRP3 signaling pathway in mice. Furthermore, PSPP alleviated Con A-induced oxidative stress by activating the PI3K/AKT/mTOR signaling pathway in mice. Additionally, PSPP demonstrated the ability to reduce inflammation and oxidative stress in RAW264.7 cells induced by Con A in vitro. ConclusionPSPP has the potential to ameliorate hepatic inflammation via the TLR4-P2X7R/NLRP3 pathway and inhibit hepatic oxidative stress through the PI3K/AKT/mTOR pathway during the progression of Con A-induced hepatic injury. The results of this study have unveiled the potential hepatoprotective properties of purple sweet potato and its medicinal value for humans. Moreover, this study serves as a valuable reference, highlighting the potential of PSPP-1 as a drug candidate for the treatment of immune liver injury.

A new synergistic hydrocolloid with superior rheology: Locust bean /xanthan gum binary solution powdered by different drying methods

In the current study, a novel gum powder containing locust bean and xanthan gums was prepared by three different drying methods of their synergistic aqueous solutions: direct cabinet drying (CD), drying after alcohol precipitation (PD), and direct freeze drying (FD). The purpose of this process is to develop ready-to-use synergistic gum powders with improved rheological properties by modifying intermolecular interactions. Significant improvements in viscosity, consistency efficiency and pseudoplastic character were observed for PD, followed by CD and FD, as well as the higher values of G′ and G″ values of PD, indicating a stronger entanglement network development between the macromolecules for PD. PD exhibited better elasticity and shear resistance compared to FD and CD in the strain sweep test. In Lissajous curves, straight line shapes were found for synergistic solutions at low strain, which continued for increasing strain values for PD and CD. However, the pronounced viscous behavior was found for FD at 500% strain. Compared to CD and FD, PD exhibited a more desirable thixotropic character. A lower zeta potential was observed in the production of synergistic gums using the PD process. Gel strength was found to be higher for PD. Therefore, PD was a more effective method to produce a ready-to-use synergistic powder with better viscoelastic properties and a more stable solution.

Characterization and anti-aging activities of polysaccharide from Rana dybowskii Guenther.

Introduction: Rana dybowskii Guenther (RDG), as a traditional Chinese medicine, has been shown to have antioxidant effects. However, studies on the anti-aging effect of RDG are still limited. Methods: In this study, we prepared polysaccharides from the skin of RDG (RDGP) by hot water extraction, alcohol precipitation, ion-exchange chromatography and gel chromatography. The proteins were removed using the Sevage method in combination with an enzymatic method. The structural features were analyzed using high-performance gel permeation chromatography, β-elimination reaction and Fourier transform infrared spectra. The anti-aging effect of RDGP was investigated by using D-Gal to establish an aging model in mice, and pathological changes in the hippocampus were observed under a microscope. Results: We obtained the crude polysaccharide DGP from the skin of RDG, with a yield of 61.8%. The free protein was then removed by the Sevage method to obtain DGPI and deproteinated by enzymatic hydrolysis combined with the Sevage method to further remove the bound protein to obtain the high-purity polysaccharide DGPII. Then, DGPIa (1.03 × 105Da) and DGPIIa (8.42 × 104Da) were obtained by gel chromatography, monosaccharide composition analysis showed that they were composed of Man, GlcA, GalNAc, Glc, Gal, Fuc with molar ratios of 1: 4.22 : 1.55: 0.18 : 8.05: 0.83 and 0.74 : 1.78: 1: 0.28: 5.37 : 0.36, respectively. The results of the β-elimination reaction indicated the presence of O-glycopeptide bonds in DGPIa. The Morris water maze test indicated that mice treated with DGPIIa exhibited a significantly shorter escape latency and increased time spent in the target quadrant as well as an increase in the number of times they traversed the platform. Pathologic damage to the hippocampus was alleviated in brain tissue stained with hematoxylin-eosin. In addition, DGPIIa enhanced the activities of SOD, CAT, and GSH-Px and inhibited the level of MDA in the serum and brain tissues of aging mice. Discussion: These results suggest that RDGP has potential as a natural antioxidant and provide useful scientific information for anti-aging research.