Ethanol Precipitation Research Articles

Structure and characterization of an extracellular polysaccharide from Paenibacillus polymyxa 88A.

Levan-type polysaccharides, produced by various organisms, are nontoxic, biocompatible, and biodegradable polymers with a wide range of biological activities. They have high potential for use in medicine, cosmetology, and industry. A large amount of levan (41.1 g L-1) was recovered by ethanol precipitation from a liquid nutrient medium of Paenibacillus polymyxa 88A that contained 15 % w/v sucrose as a carbon source. The levan was fractionated by gel-permeation and anion-exchange chromatography and was analyzed by DRIFT and NMR spectroscopy. It was found that levan was represented by slightly branched chains composed of β-(2→6)-Fruf residues. The average molecular mass of the levan was about 1.9 MDa. When shear stress was applied at different temperatures, aqueous levan solutions showed pseudoplastic behavior. As found by SEM, a freeze-dried powdered levan sample had a microporous structure. The levan had excellent emulsifying activity toward sunflower oil, forming an emulsion with long-term stability. Analysis of the antioxidant activity of the levan showed a higher, dose-dependent activity toward ABTS, as compared with that toward DPPH. Finally, the bioactivity of the levan was examined by MTT assay by using human cervical carcinoma (HeLa) cells.

Physicochemical Properties and Biological Activities of Polysaccharides from Panax Notoginseng Separated by Fractional Precipitation.

The dried root of Panax notoginseng is a medicinal and food ingredient. P. notoginseng polysaccharides (PNPs) have physicochemical properties, which have not been fully elucidated. This study aimed to identify a method to separate the PNP fractions and investigate their activities. PNPs were prepared from roots by hot water extraction, deproteinization, and decolorization. PNP20, PNP40, and PNP60 fractions were isolated through stepwise ethanol precipitation at 20 %, 40 %, and 60 % concentrations, respectively. The three polysaccharide fractions were characterized using chromatography, spectroscopy, and thermogravimetric analysis, and their moisture retention, antioxidant, and tyrosinase-inhibition properties were evaluated. Monosaccharide composition analysis showed that the three PNPs contained mannose (Man), galacturonic acid (GalA), glucose (Glc), galactose (Gal), and arabinose (Ara) in different molar ratios. HPGPC analysis demonstrated that the polysaccharides precipitated with higher ethanol concentrations had lower molecular weights (Mw). Furthermore, all PNPs had distinct moisturizing and hygroscopic properties and antioxidant activities, with PNP60 showing better antioxidant properties and a competitive mixture of hygroscopic properties and tyrosinase inhibition. The chemical composition and structural characteristics of PNPs could affect their functional attributes. PNP60 has the potential to be a moisturizer and antioxidant and could be used in the development of cosmetic ingredients.

Total RNA Extraction from Starchy Rice Endosperm.

RNA extraction is a critical step in gene expression analysis. While numerous RNA extraction kits are commercially available, most kits cannot be utilized for RNA extraction from rice endosperm that contains abundant starch. Here, I describe a three-step RNA extraction from rice starchy endosperm. The process involves phenol/chloroform extraction, ethanol precipitation, and lithium chloride precipitation. This method yields an abundant and high-quality total RNA even from a single starchy endosperm.

Structural Characteristics and Immunological Function of a New Non-Starch Polysaccharide from Red Sprout Taro

Taro is a tuber crop that is used for nutritional and medicinal purposes due to its abundance of non-starch polysaccharides (NSPs). Red Sprout taro is a local variety in Southern China, but the characteristics and bioactivities of its NSPs are currently unknown. In this study, NSPs were isolated from the corms of Red Sprout taro using hot-water extraction, ion-exchange chromatography, and ethanol precipitation; their molecular weight, monosaccharide composition, structural formulae, and immunomodulatory effects were examined. A novel NSP named Colocasia esculenta polysaccharide 1 (CEP1) was purified and characterized and was shown to mainly consist of glucose (60.49%) and galactose (25.92%) and have a molecular weight of 4556.272 kDa. The backbone of CEP1 consisted primarily of →4)-α-D-Glcp-(1→, →4,6)-β-D-Galp-(1→, and →3)-β-D-Galp-(1→ residues, with a branch consisting of the β-D-Glcp-(1→ residue. In addition, 25–400 µg/mL CEP1 was shown to have immunomodulatory effects on RAW264.7 macrophages. CEP1 not only increased cell viability, phagocytic capacity, inducible nitric oxide synthase secretion, and nitric oxide generation in RAW264.7 cells, but it also activated M1 and M2 macrophages to generate tumor necrosis factor α, interleukin 6, transforming growth factor β, and interleukin 10. These findings could lead to the use of CEP1 from Red Sprout taro as a possible immunomodulatory polysaccharide in functional foods.

A polysaccharide from Periplaneta americana promotes macrophage M2 polarization, exhibiting anti-inflammatory and wound-healing activities

A miscellaneous polysaccharide, PAP55–3-1, with a molecular weight of 23.03 kDa, was isolated from Periplaneta americana through extraction with dilute alkali solution, ethanol precipitation, and column chromatography purification. Structural analysis shows that PAP55–3-1 is mainly composed of five monosaccharides: galactosamine hydrochloride, glucosamine hydrochloride, galactose, glucose and mannose. Its main glycosidic bonds are: Manp-(1→, Galp-(1→, →3)-Galp-(1→, →3,6)-Manp-(1→, →2,6)-Manp-(1→, →6)-Manp-(1→, →4)-Galp-(1→, →6-Glcp-(1→, →6)-Galp-(1→, →2)-Manp-(1 →, →3,4)-Glcp-(1→, →3,6)-Galp-(1→. In vitro experiments demonstrated that PAP55–3-1 can effectively inhibit reactive oxygen species (ROS) and O2− production following H2O2−induction. After H2O2−induction, HIF-1α (hypoxia-inducible factor) was translocated in mitochondria PAP55–3-1 increased localization of HIF-1α was located on mitochondria to maintain the stability of mitochondrial function stability, thereby effectively inhibiting H2O2-induced mitochondrial oxidative damage. Additionally, PAP55–3-1 inhibited the M1 polarization of macrophages stimulated by H2O2 and promoted the phenotype polarization of macrophages from M1 to M2, displaying anti-inflammatory and pro-repair properties. In vivo experimental results indicated that PAP55–3-1 promoted wound healing in mice. Immunohistochemical experiments revealed a reduction in CD68 expression and increase in CD206 expression in both positive and the high-dose polysaccharide group control group. This further demonstrated that PAP55–3-1 promotes the phenotype polarization of macrophages from M1 to M2, exerting anti-inflammatory and wound-healing activities.

Effect of H2O2/ascorbic acid degradation and gradient ethanol precipitation on the physicochemical properties and biological activities of pectin polysaccharides from Satsuma Mandarin

In this work, three degraded polysaccharides (DMPP-40, DMPP-60, DMPP-80) were successfully obtained by H2O2/ascorbic acid degradation and gradient ethanol precipitation from Satsuma mandarin peel pectin (MPP), and their physicochemical properties, antioxidant and prebiotic activities were investigated. The molecular weight of MPP, DMPP-40, DMPP-60, DMPP-80 were determined to be 336.83 ± 10.57, 18.93 ± 0.54, 26.07 ± 0.83 and 8.71 ± 0.27 kDa, respectively. The ethanol concentration significantly affected the physicochemical properties of DMPPs. DMPP-60 showed the highest yield (69.07 %) and uronic acid content (64.85 %), DMPP-80 showed the lowest molecular weight (8.71 kDa), and the composition and proportion of monosaccharides of DMPPs were significantly different. Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance spectroscopy (1H NMR) confirmed that DMPPs exhibited similar functional groups, while X-ray diffraction (XRD) indicated that DMPP-40 possessed some crystallographic sequences. Scanning electron microscopy (SEM) images directly verified the fragmented structure and reduced surface area of DMPPs. Besides, the H2O2/ascorbic acid treatment could obviously reduce the apparent viscosity and thermal stability of MPP. Meanwhile, the results of bioactivity assay showed that DMPPs possessed better antioxidant activity and probiotics pro-proliferative effects compared with MPP. DMPP-80 could significantly inhibit lipopolysaccharides (LPS)-stimulated production of inflammatory factors (including nitric oxide (NO), interleukin (IL)-6, tumor necrosis factor (TNF)-α and interleukin (IL)-1β) in RAW264.7 cells. Results suggest that the H2O2/ascorbic acid combined with gradient ethanol precipitation has potential applications in degradation and separation of MPP to improve its biological activities.

Optimization of manufacturing process for serotype 14 pneumococcal capsular polysaccharide

Streptococcus pneumoniae is a pathogenic bacterium that causes infections such as pneumonia, meningitis, otitis media, and bacteremia. The prevention of pneumococcal disease by vaccination has become more urgent due to increased antibiotic resistance. Pneumococcal capsular polysaccharides (CPS) are effective vaccine antigens that stimulate the host to produce protective antibodies. S. pneumoniae serotype 14 is one of most prevalent types in Latin America and across the world. However, the yield of S. pneumoniae serotype 14 CPS from existing fermentation processes remains low and requires improvement. In this study, various aspects of the fermentation process were optimized to improve pneumococcal growth and polysaccharide productivity, including feed medium, cultivation gas environment, fermentation pH, and temperature. A simplified purification method was also developed to obtain pure CPS, including ultrafiltration, acid and ethanol precipitation, diafiltration, and lyophilization. These fermentation optimizations significantly enhanced the optical density of pneumococcal bacterial cultures and increased fermentation yields to 2.4–2.6 g/L—significantly higher than previously achieved. Furthermore, the test results of pure CPS could meet the requirements in the European Pharmacopoeia (11th edition). These optimizations provide valuable insights into the nutritional requirements and impact of varying fermentation process parameters on pneumococcal growth and CPS productivity, thus contributing to the development of a more efficient and cost-effective method for the production of pneumococcal CPS—essential for manufacturing vaccines against pneumococcal infections.

Study on Preparation and Properties of Pickering Emulsions Prepared by Carboxymethyl Starch‐Xanthan Gum Nanoparticles

AbstractIn this work, carboxymethyl starch‐xanthan gum nanoparticles (CXN) with different ratios are prepared by ethanol precipitation method. The effects of different ratios, NaCl content, and pH on the particle size and zeta potential of the nanoparticles are investigated, as well as the rheological properties and stability of the CXN emulsions. The results show that with the increase of xanthan gum (XG), the particle size, and zeta potential of the nanoparticles decrease and the prepared emulsions have higher shear stress and viscosity, as well as greater thermal and storage stability. Changes in pH and NaCl concentration significantly affect the particle size and zeta potential values of the nanoparticles. Excess acids or bases increase the viscosity of the emulsions. At low salt concentrations (0.05 M), the emulsion viscosity increases significantly, after which the emulsion viscosity decreases with increasing salt concentration. Neutral pH conditions at room temperature are not conducive to the stability of the emulsions. At high temperatures (100 °C), high salt concentrations, excessive acids or bases can destabilize emulsions. This study provides a new way to reduce the particle size of nanoparticles and a feasible strategy for developing long‐term stable Pickering emulsion.

Anti-Melanogenic Effects of a Polysaccharide Isolated from Undaria pinnatifida Sporophyll Extracts.

Undaria pinnatifida is a temperate brown alga known to exert free radical-scavenging and anti-inflammatory effects. In this study, we investigated the skin-whitening effects of U. pinnatifida sporophyll extracts (UPEs) in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 melanoma cells. The crude polysaccharide fraction (UPF) was obtained via ethanol precipitation. Four polysaccharide fractions (UPF1-4) were isolated and purified using ion-exchange column chromatography, and their anti-melanogenic activity was evaluated. UPF3 exhibited the highest anti-melanogenic activity, showing the highest sulfate (39.79%), fucose (143 μg/mg), and galactose (208 μg/mg) contents. UPF3 significantly inhibited intracellular tyrosinase activity in B16F10 cells. We also evaluated the melanogenic signaling pathway to determine the mechanism of action of UPF3 in melanongenesis. UPF3 reduced the expression of tyrosinase-related protein-1 (TRP-1), tyrosinase-related protein-2 (TRP-2), and tyrosinase, which play important roles in melanin production. Therefore, UPF3 has high potential for use in skin-whitening functional pharmaceuticals and cosmetics.

Decoding of novel umami peptides from corn fermented powder and its mechanism via multisensory techniques, virtual screening, and molecular simulation approaches

This study aimed to identify umami peptides in corn fermented powder (CFP) and investigate their umami enhancing effect. Ultrafiltration and ethanol precipitation was used to separate the umami peptides in CFP. Dynamic sensory evaluations were used to identify the peptide fraction with the intense umami taste, and the peptides in the fraction were identified by nano-liquid chromatography-tandem mass spectrometry. Subsequently, ten umami-enhancing peptide candidates were screened using an integrated virtual screening strategy. Molecular docking revealed that Ser382, Ser104, Leu334, Glu338 and Glu148 of the T1R1 and T1R3 taste receptors are important amino acid residues for binding of the ten umami peptides. Three umami peptides (VDW, WGDDP, and WPAGE) exhibited the stronger binding affinity with the umami receptors. Moreover, molecular dynamics simulation revealed that the T1R1/T1R3 formed stable complexes with the three umami peptides during the simulation. Sensory evaluation indicated that the three peptides exhibited diverse taste characteristics (detection thresholds:0.0315–0.0625 mg/mL). The sigmoid curve analysis further confirmed peptides were identified as synergistically (VDW and WGDDP) or additively (WPAGE) enhancing the umami of 3 mg/mL MSG solution. This study uncovers the mechanism of umami-peptide-driven taste in fermented corn products.

Analysis of sediment re-formation factors after ginseng beverage clarification based on XGBoost machine learning algorithm

The aim of this study was to explore the sediment re-formation factors of ginseng beverages subjected to four clarification ways (11 subgroups) including the ethanol precipitation, enzymatic treatment, clarifier clarification, and Hollow Fiber Column (HFC) methods, based on the Extreme Gradient Boosting (XGBoost) model. The results showed that the clarity of the ginseng beverages was significantly improved by all the clarification treatments, but still formed sediment after storage. HFC method exhibited the highest transmittance, the least sediment, and stronger antioxidant activity in the clarification treatment groups. According to the results of chemical composition analyses and partition coefficients, carbohydrates, saponins, proteins and metal elements were involved in varying degrees in the re-formation of the sediments in ginseng beverage after clarification. Based on the above data, the XGBoost model predicted that protein, Rd, Na, K, and total saponins were the five most important chemical components affecting the sediment re-formation in ginseng beverages.

Distinct prebiotic effects of polysaccharide fractions from Polygonatum kingianum on gut microbiota

This study investigated the physicochemical properties, digestive stability, and in vitro fermentation behavior of Polygonatum kingianum polysaccharide (PKP) fractions (PKP60, PKP70, PKP80) obtained through graded ethanol precipitation. High-performance gel permeation chromatography revealed significant molecular weight differences among the fractions, while reverse-phase high-performance liquid chromatography indicated consistent monosaccharide types with variations in their proportions. Uronic acid analysis confirmed that all polysaccharide fractions met the criteria for neutral polysaccharides. Congo red staining confirmed the presence of a triple-helix structure in all PKP fractions. Comprehensive analysis demonstrated that these fractions remained stable during in vitro digestion, as evidenced by consistent molecular weights and total carbohydrate content, with no significant production of free monosaccharides or reducing sugars. All PKP fractions were fermented by gut microbiota, resulting in the production of short-chain fatty acids. Beta diversity and structural analyses of gut microbiota revealed distinct modulatory effects associated with each PKP fraction. The PKP fractions promoted probiotic growth, especially PKP70, which significantly enhanced Bifidobacterium proliferation, indicating strong prebiotic potential. These findings underscore the importance of isolation and purification methods in determining the functionality and gut microbiota-modulating effects of plant-derived polysaccharides, emphasizing the need for in-depth research that extends beyond merely evaluating their source.

In vitro and in vivo effects of pectin‐type polysaccharides isolated from crabapples (Malus prunifolia) on inflammatory colitis models

SummaryThe aim of this study was to investigate the anti‐inflammatory effects of polysaccharides extracted from crabapples (Malus prunifolia; MP) in vitro and in vivo. Crabapples were subjected to hot water extraction to obtain MP‐HW, which was further separated into low (MP‐L) and high molecular weight (MW) (crude polysaccharide; MP‐CP) fractions by ethanol precipitation. Based on MW distribution, general composition, component sugars, and Fourier transform infrared spectroscopy, MP‐CP was identified as a pectin‐type polysaccharide rich in galacturonic acid with an MW of 106.2 kDa. MP‐CP inhibited the production and mRNA expression of proinflammatory factors in an IL‐1β‐treated intestinal epithelial cell model. In addition, MP‐CP significantly alleviated the body weight, disease activity index score, colon length, and spleen weight in mice with dextran sulfate sodium (DSS)‐induced colitis. The MP‐CP‐treated group showed effective regulation of the expression of pro‐ and anti‐inflammatory factors and immunoglobulin A than the DSS‐treated group. Haematoxylin and eosin and Alcian blue staining revealed that MP‐CP alleviated inflammatory manifestations and mucin expression in the intestinal lumen. Collectively, pectin‐type polysaccharides isolated from crabapples may be used as a new functional food and pharmaceutical agent for the prevention and treatment of inflammatory colitis.

Polysaccharides from pineapple peel: Structural characterization, film-forming properties and its effect on strawberry preservation

The growing demand for food safety has stimulated the development of new environmentally friendly food packaging. It is the development trend of food packaging in recent years by using natural polysaccharides as carriers and adding bioactive ingredients extracted from plants to prepare multifunctional films with antioxidant, antimicrobial and biodegradable properties. Herein, three polysaccharide components (PPE40, PPE60, and PPE80) from pineapple peel were extracted by ultrasound-assisted hot water extraction combined with gradient ethanol precipitation method, which all showed a certain scavenging activities against DPPH, ABTS, and hydroxyl radical. Then, the composite films were prepared by adding PPE40, PPE60 and PPE80 to chitosan. The results of SEM, FT-IR and XRD analysis showed that PPE40, PPE60 and PPE80 could interact with chitosan matrix. Furthermore, the addition of PPE40, PPE60, and PPE80 could improve the mechanical properties of the films, and promote the antibacterial activity of the films against B. subtilis, S. aureus and E. coli. Finally, the application of the composite films to strawberries showed that the addition of PPE40, PPE60 and PPE80 could delay the rapid decay of strawberries during storage. The results of this study showed that pineapple polysaccharides have a potential to be applied in the field of food packaging.

Structural elucidation of a capsular polysaccharide from Bacteroides uniformis and its ameliorative impact on DSS-induced colitis in mice

Capsular polysaccharides derived from Bacteroides species have emerged as potential mitigators of intestinal inflammation in murine models. However, research on capsular polysaccharides from B. uniformis, a Bacteroides species with reduced abundance in colons of patients with ulcerative colitis, remains scarce. In this study, we extracted a neutral polysaccharide component from B. uniformis ATCC8492, termed BUCPS1B, using ultrasonic disruption, ethanol precipitation, and anion exchange chromatography. Structural characterization revealed BUCPS1B as a water-soluble polysaccharide with an α-1,4-glucan main chain adorned with minor substituent sugar residues. BUCPS1B alleviated intestinal inflammation in a mouse model of colitis and induced polarization of macrophages into M2-type. Furthermore, BUCPS1B modulated the gut microbiota composition, increased the abundance of the probiotic Akkermansia muciniphila and altered the gut metabolic profile to promote phenylalanine and short chain fatty acids metabolism. BUCPS1B is therefore a promising candidate to prevent inflammation and augment intestinal health.

A novel glycopeptide from mountain-cultivated ginseng residue protects type 2 diabetic symptoms-induced heart failure

Ethnopharmacological relevanceMountain-cultivated Panax ginseng C.A.Mey. (MCG) with high market price and various properties was valuable special local product in Northeast of Asia. MCG has been historically used to mitigate heart failure (HF) for thousand years, HF is a clinical manifestation of deficiency of “heart-qi” in traditional Chinese medicine. However, there was little report focus on the activities of extracted residue of MCG. Aim of the studyA novel glycopeptide (APMCG-1) was isolated from step ethanol precipitations of alkaline protease-assisted extract from MCG residue. Materials and methodsThe molecular weight and subunit structure of APMCG-1 were determined by FT-IR, HPLC and GPC technologies, as well as the H9c2 cells, Tg (kdrl:EGFP) zebrafish were performed to evaluated the protective effect of APMCG-1. ResultsAPMCG-1 was identified as a glycopeptide containing seven monosaccharides and seven amino acids via O-lined bonds. Further, in vitro, APMCG-1 significantly decreased reactive oxygen species production and lactate dehydrogenase contents in palmitic acid (PA)-induced H9c2 cells. APMCG-1 also attenuated endoplasmic reticulum stress and mitochondria-mediated apoptosis in H9c2 cells via the PI3K/AKT signaling pathway. More importantly, APMCG-1 reduced the blood glucose, lipid contents, the levels of heart injury, oxidative stress and inflammation of 5 days post fertilization Tg (kdrl:EGFP) zebrafish with type 2 diabetic symptoms in vivo. ConclusionsAPMCG-1 protects PA-induced H9c2 cells while reducing cardiac dysfunction in zebrafish with type 2 diabetic symptoms. The present study provides a new insight into the development of natural glycopeptides as heart-related drug therapies.

Hydrothermal extraction of ulvans from Ulva spp. in a biorefinery approach

A simple cascade process based on the hydrothermal fractionation of Ulva spp. biomass was proposed. Considering the overall extraction yields (50 %), ulvan recovery (23 %), and ulvan composition, structural, mechanical and cytotoxic properties, the selected optimal final heating temperature was 160 °C. Ethanol precipitation provided the highest ulvan recovery yields but choline chloride precipitated ulvans showed stronger mechanical properties, G´ moduli 1.5·104 Pa and 3·104 Pa for ethanol and for choline chloride, respectively. Both products were safe on NCTC 929 mouse fibroblasts and after a cooling stage, formed films without requiring any additives. From the ulvan-free liquid fraction, one product with 43 % (wt, d.b.) phenolics and moderate antiradical properties and a byproduct containing nutrients and minerals were separated. The methane potential of the corresponding residual solids was influenced by the hydrothermal heating temperature and was doubled compared to than for the untreated seaweed biomass (60 mL/g VS). This scheme could be also applied to the wet algal biomass, in a chemical free alternative to provide ready to use ulvan biopolymers, bioactives, nutrients, salts and biogas, conforming a biorefinery approach.

Production and partial purification of an innovative heat resistant α-keratinase with some remarkable medical and industrial applications

Background and objectives Keratinase has ultimate practical importance in industry, medicine, food industry and waste management fields. Their applications in wool and silk are as good cleaners and in leather industry as the best ever green dehairing agents providing high leather quality, as well they are used as crucial components of sophisticated detergents. Their medical prospective applications are in prion degradation and human callus removal. They convert keratinaceous wastes to valuable products saving the environment from hard keratin waste pollution. Bacteria, fungi and actinomycetes are effective keratinase producers and they are considered the most suitable sources. This study aims to formulate the production medium and to pinpoint the proper physiological conditions for the potent microorganism producing an efficient α-keratinase enzyme. The partial purification of the crude enzyme was successfully performed. The effect of the reaction temperature on both the crude and the partially purified enzyme (PPE) was duly studied with the thermostability of PPE. Some important applications have been implemented on PPE and these include leather dehairing, cloth stain removal, and topical treatment of human callus. Materials and Methods Thirteen recommended microbial strains were screened for effective and applicable α-keratinase productivity. Optimization of the cultural conditions for extracellular enzyme production and also the partial purification of the crude enzyme by ammonium sulphate salting out or by ethanol or acetone precipitation were carried out. The effect of reaction temperature on the enzyme and its thermostability were studied. Finally, the efficiency of the PPE on leather dehairing, stain removal, and human callus treatment was explored. Results and conclusion Among the 13 organisms screened, the fungal strain Trichoderma polysporum HZ-31 was the most potent producer of an influencial α-keratinase. The maximum α-keratinase activity of 58.2 UmL−1 was obtained by the previous-mentioned strain after 5-days fermentation medium containing (%, w/v): whole chicken feathers 0.5, glucose 0.2, peptone 0.5, yeast extract 0.5, K2HPO4 0.1, KH2PO4 0.3, CaCl2 0.1, MgSO4 0.1, and pH 7.0. Acetone fractionation of the crude keratinase was the most proper and offered the most promising keratinase fraction PPE at 80–90% acetone. This fraction had high thermostability and was kept at 55°C for more than 98% of its original activity after 60 min heating and this temperature (55°C) was also the optimum for 2 h enzymatic reaction. Conclusively, the present study succeeded in the achievement of a constitutive extracellular alkaline α-keratinase, which successfully proceeded to complete leather unhairing after 12–16 h at 37°C, afforded high performance to cloth blood stain removal with Arial detergent after 2 h at 50°C and complete degradation of the human callus after 4 h at 50°C.

Separation and purification of bovine nasal cartilage-derived chondroitin sulfate and evaluation of its binding to bovine serum albumin

This study employs an optimized and environmentally friendly method to extract and purify chondroitin sulfate (CS) from bovine nasal cartilage using enzymatic hydrolysis, ethanol precipitation, and DEAE Sepharose Fast Flow column chromatography. The extracted CS, representing 44.67 % ± 0.0016 of the cartilage, has a molecular weight of 7.62 kDa. Characterization through UV, FT-IR, NMR spectroscopy, and 2-aminoacridone derivatization HPLC revealed a high content of sulfated disaccharides, particularly ΔDi4S (73.59 %) and ΔDi6S (20.61 %). Interaction studies with bovine serum albumin (BSA) using fluorescence spectroscopy and molecular docking confirmed a high-affinity, static quenching interaction with a single binding site, primarily mediated by van der Waals forces and hydrogen bonding. The interaction did not significantly alter the polarity or hydrophobicity of BSA aromatic amino acids. These findings provide a strong foundation for exploring the application of CS in tissue engineering and drug delivery systems, leveraging its unique interaction with BSA for targeted delivery and enhanced efficacy.

Extraction of basil seed gum: Optimization and functional properties

Basil seed gum (BSG), a natural vegetable gum, was extracted by ethanol precipitation method using basil seeds as raw material. Based on the single factor experiments, Box-Behnken response surface test was performed to optimize the extraction conditions, and the physicochemical and functional properties of the extracted basil seed gum, as well as its microstructure, were analyzed in detail. The results showed that the optimal extraction conditions of basil seed gum were as follows: liquid-to-feed ratio of 62:1, temperature of 50 °C, extraction time of 29.50 min, and pH = 8.1, with yield of 11.28 %. BSG was mainly composed of total sugars (93.09 %), uronic acid (18.83 %), proteins (2.77 %), and ash (4.35 %), and its monosaccharides included D-(+)-anhydrous glucose (58.26 %), D-galactose (21.40 %), D-mannose (11.96 %), D-(+)-galacturonic acid (7.82 %), D-arabinose (0.28 %), D-(+)-xylose (0.28 %), and L-rhamnose (0.002 %). Comparative analysis of the functional properties of basil seed gum and three commercial vegetable gums (linseed gum, caraway seed gum, guar gum) showed that basil seed gum had excellent water retention properties, and its water absorption and water holding properties were much better than those of three commercial gums. Rheological characterization showed that BSG was a pseudoplastic fluid with high zero-shear viscosity. In addition, the basil seed gum powder showed an irregular flaky fibrous structure under microscope and was a semi-crystalline polymer. In view of the high aqueous absorption and retention properties of basil seed gum, it has promising applications in food, cosmetic and medical devices.