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. 2023 Feb 21;13(3):93. doi: 10.1007/s13205-023-03506-1

Xylans extracted from branches and leaves of Protium puncticulatum: antioxidant, cytotoxic, immunomodulatory, anticoagulant, antitumor, prebiotic activities and their structural characterization

Iranildo José da Cruz Filho 1, Thammyris Pires de Souza 1, Carolina Ávila dos Anjos Santos 1, Maria Augusta de Morais Araújo 1, Josué Filipe de Oliveira Moraes Miranda 1, Maria Emanuelle de Oliveira Queirós 1, Denilson José Nogueira Correia Filho 1, Alice da Conceição Alves de Lima 1, Diego Santa Clara Marques 1, Maria do Carmo Alves de Lima 1,✉
PMCID: PMC9944590  PMID: 36845077

Abstract

This work aimed to isolate and characterize xylans from branches and leaves of Protium puncticulatum, in addition to evaluating its in vitro biological and prebiotic potential. The results showed that the chemical structure of the obtained polysaccharides is similar being classified as homoxylans. The xylans presented an amorphous structure, in addition to being thermally stable and presenting a molecular weight close to 36 g/mol. With regard to biological activities, it was observed that xylans were able to promote low antioxidant activity (< 50%) in the different assays evaluated. The xylans also showed no toxicity against normal cells, in addition to being able to stimulate cells of the immune system and showing promise as anticoagulant agents. In addition to presenting promising antitumor activity in vitro. In assays of emulsifying activity, xylans were able to emulsify lipids in percentages below 50%. Regarding in vitro prebiotic activity, xylans were able to stimulate and promote the growth of different probiotics. Therefore, this study, in addition to being a pioneer, contributes to the application of these polysaccharides in the biomedical and food areas.

Supplementary Information

The online version contains supplementary material available at 10.1007/s13205-023-03506-1.

Keywords: Amazon rainforest, Biological activities, Polysaccharides, Xylans

Introduction

Xylans are the most abundant type of hemicellulose in nature (Deutschmann et al. 2012). With a complex macromolecular structure, this polysaccharide has a main chain composed of xylose residues joined by β-1,4-glycosidic bonds (Zhang et al. 2022). In addition to the main chain, xylans can have dimeric and trimeric side chains, formed by different monosaccharides and functional groups (Naidu et al. 2018). The chemical structure of these polysaccharides may vary according to the plant source and the production method used (Jana 2021).

Xylans have been the subject of several studies for their ability to promote various biological activities, among which we can mention: antioxidants (Bhanja et al. 2022), anti-tumor (Qian et al. 2022), immunomodulators (Pynam et al. 2019), emulsifier (Wang et al. 2022) and functional foods (Chen et al. 2019). Due to the versatility of applications and biological properties, it is estimated that products obtained from xylans will generate revenue of US$ 130 million by 2025 (Jana 2021).

In this perspective, the search for new sustainable sources of obtaining xylans has become a promising alternative for the use of these polysaccharides as raw material. This use is directly related to availability, commercial viability and plant growth. Among the different sources of obtaining, we highlight here Protium puncticulatum species, a tree from the Amazon rainforest that is very important for the timber sector. Obtained as reforestation wood, this tree has a basic density of 0.55–0.60 g/cm3, with color ranging from grayish brown to light reddish brown, sapwood and heartwood is not very distinct, indistinct growth rings, straight grain, medium to fine texture, strong shine and has good workability, being used in woodworking, boxing and construction in general (Iwakiri et al. 2016).

Therefore, this work aimed to obtain two xylans extracted from branches and leaves of Protium puncticulatum. The branches and leaves were chosen because they are considered promising residues from timber companies. These are obtained in large quantities during the process of pruning the trees and during the obtaining of the wood. Due to its abundance, branches and leaves can be used sustainable as raw material for the production of energy, fertilizers and various bio-products. The novelty of this study is the pioneering application of xylans in different in vitro biological assays and structural characterization. All these tests were carried out to evaluate these polysaccharides as a raw material. This is to obtain products with high added value, which can be used in various industrial and pharmaceutical sectors.

Materials and methods

Plant material: Protium puncticulatum

The branches and leaves used in this work were kindly provided by the company Mil Madeiras Preciosas Ltd located in the municipality of Itacoatiara in the State of Amazonas, Brazil. The species was registered in SisGen, no. A36E658. The branches and leaves were dried at 60 ºC for 48 h in an oven, then ground in a knife mill and sieved through an 80 mesh.

Chemical composition analysis

The contents of cellulose, hemicellulose, lignin, extractives and ash present in the branches and leaves of Protium puncticulatum were determined using the methodology proposed by Araujo et al. (2022).

Process of extraction and obtaining of xylans

The xylans were obtained according to Khaire et al. (2021) under conditions previously optimized by the authors. Initially, delignification was performed by adding 500 mL of a solution containing 35 g of sodium chlorite (NaClO2) and 2.5 mL of glacial acetic acid to 50 g of branches or leaves. The process occurred in a ratio of 1:10 (w/v) in a 1L bottle. Then the system was incubated at 60 °C for 3 h.

After this period, the solid was filtered with distilled water until pH 7.0. A 346 mL solution containing 10% NaOH and 1% H3BO4 was mixed with the delignified material (34.6 g) and stirred at 200 rpm for 3 h at 60 °C. The suspension was filtered and the xylan was precipitated with 3 volumes of cold ethanol (95%) containing 10% acetic acid and incubated at 4 °C for 12 h.

The xylan obtained was resuspended in 200 mL of water and dialyzed in 4 L of ultrapure water. Finally, the xylan was lyophilized. The yield was calculated considering the amount of hemicellulose in the raw material according to Eq. 1.

Yield%=Xylan mass obtained (g)Total mass of dry material (g)×100% 1

Thus, two xylans were obtained from the branches (PPB) and leaves (PPL) of Protium puncticulatum.

Physicochemical characterization

Scanning electron microscopy (SEM)

The morphology of xylans, scanning electron microscopy technique was used. The analyses were performed according to Khaire et al. (2021) with modifications. The samples were previously coated with 20 nm gold in a metallizer JFC-1100 and analyzed in a scanning electron microscope JEOL T-200.

Analysis of the composition of monosaccharides

The xylans (2 mg) were subjected to acid hydrolysis under the conditions defined by Yan et al. (2018) with modifications. The acid hydrolysis was performed at 121 °C in a thermostatic bath, using 2.5 mL of trifluoroacetic acid (2 M) for 6 h. The identification and quantification of the compounds present in the hydrolysed was performed using HPLC, according to the methodology proposed by Arruda et al. (2021). For this purpose, the Aminex HPX87H column was used, temperature of 60 °C, with mobile phase: H2SO4 5 mM, flow rate of 0.6 mL/min and refractive index detector for identification and quantification.

Elementary analysis

The percentages of carbon, hydrogen and nitrogen present in the xylans (20 mg) were determined in an elemental analyzer. The oxygen content was obtained by difference in the percentages of carbon, nitrogen and hydrogen.

Fourier transform attenuated total reflection spectroscopy analysis

Approximately 100 mg of xylan was pressed to form a 12 mm diameter, 1 mm thick disc. The spectra were obtained in a Perkin Elmer spectrophotometer (FT-IR Spectrum Two – UART Two) in the region from 600 to 4000 cm−1.

Characterization by nuclear magnetic resonance

The xylans (20 mg) were dissolved in deuterated water and analyzed in a BRUKER Avance 400 MHz spectrometer at 20 °C.

Thermal degradation

Thermogravimetry (TGA), differential thermogravimetry (DTG) and Differential Scanning Calorimetric (DSC) were performed according to the methodology proposed by Khaire et al. (2021) with modifications. The tests were carried out in a nitrogen atmosphere with a flow rate of 20 mL/min to avoid thermal oxidative degradation from 5.1 mg of the powder sample, with a heating rate of 10 °C/min. The temperature range was from 30 to 600 °C (TGA/DTG). The initial mass of the samples was 4.0 mg. DSC temperature range from 0 to 300 °C. The experiments were performed in triplicate.

X-ray diffraction (XRD)

XRD analyzes were performed in a diffractometer (XRD-6000/Shimadzu) was used. The conditions applied were: 40 kV, angular range 4º to 60º (Bragg angle–2θ), angular variation 0.05º and counting time of 1 s.

Determination of total phenolic compounds

The content of total phenols was determined by the method described by Ebringerová et al. (2008) with few modifications. Initially, a volume of 2.0 mL of Folin–Ciocalteu solution (1:10) was added to 4.0 mL of a xylan solution (1 mg/mL) for 3.0 min. After incubation, 1.6 mL of Na2CO3 solution (7.5%) was added and then the system was again incubated in the dark at 25 °C for 120 min. The determination of the phenolic content was performed through an analytical curve of absorbance (765 nm) as a function of the concentration of gallic acid (0 to 500 μg/mL).

Determination of molecular mass by gel permeation chromatography (GPC)

The determination of molecular mass was carried out according to the methodology proposed by Zhang et al. (2022) with modifications. GPC analyzes of the various samples were performed on a PL-GPC 110 system with two Plgel 10 µm 300 × 7.5 mm columns protected by a Plgel 10 µm precolumn. The columns, pre-column, injection system and detector (refractive index) were maintained at a constant temperature of 70 °C. The eluent used was N,N-dimethylacetamide (DMA) with 0.1 M LiCl at a flow rate of 0.9 mL/min. Approximately 2 mg of xylans was dissolved in 70 µL of (DMA) with 10% LiCl. Subsequently, the samples were diluted with DMA to an approximate concentration of 0.4%. Calibration of the GPC columns was done with pullulan standards in a range of 800–10,000 Da.

Cytotoxicity assays

Cytotoxicity in normal and tumor cells

Cytotoxicity analyzes were performed using the MTT technique, described by Bhanja et al. (2022) and Albuquerque Nerys et al. (2022) with modifications. The strains evaluated were: macrophages (J774), fibroblasts (V79) and Vero cells (normal cells); MCF-7, T-47D, Jurkat, DU145 and HepG2 (tumor cells). The xylans were dissolved in ultrapure water at an initial concentration of 400 μg/mL. Then the solutions were then filtered through 0.22 µm membranes.

Initially, cells were seeded in 96-well plates containing RPMI medium added with phenol red and supplemented with 10% fetal bovine serum and 1% penicillin:streptomycin solution, at the concentration of 1 × 106 cell/well and incubated in an atmosphere at 5% CO2 and 37 ºC. After 24 h, the medium was removed and the cells were presented in the presence of different xylans (6.25–200 μg/mL) for a 72-h period. After this interval, a volume of 10 μL of MTT (5 mg/mL) was added and the plates were incubated for 3 h in an oven at CO2 and 37ºC. Then the formazan crystals formed when solubilized in DMSO an absorbance of each assay was determined spectrophotometrically at 540 nm. Wells containing RPMI and MTT medium were used as experimental controls. Assays were performed in biological triplicate and technical quadruplicate.

The standards amsacrine, asulacrine and doxorubicin (diluted in 1% DMSO at the same concentrations as the xylans). The IC50 (concentration that inhibits growth by 50%) was determined non-linearly. The experimental control consisted only of cells and culture medium.

In vitro hemolytic activity

The hemolytic activity assay was performed according to Kumar et al. (2021) and Albuquerque Nerys et al. (2022) with few modifications. The blood used for the experiment was collected from adult mice. Erythrocytes were isolated by centrifugation (1500 rpm, 10 min at 4 °C) and washed three times with phosphate buffered saline (PBS; pH 7.4). Each tube received 1.1 mL of erythrocyte suspension (1%) and 0.4 mL of various concentrations of xylans (6.25–200 µg/mL). Controls were saline only (negative) and Triton X100 (positive). After 60 min of incubation the cells were centrifuged and the absorbance (ABS) of the supernatant was recorded at 540 nm. Assays were performed in biological triplicate and technical quadruplicate. Hemolytic activity was calculated using Eq. 2.

Hemolysis(%)=ABS Xylan - ABS negative controlABS Triton X control - ABS negative control×100% 2

In vitro anticoagulant activity assays

Blood plasma was obtained from healthy adult mice. Initially, plasma was separated from blood cells by centrifugation at 3000 rpm at 4 °C for 20 min then added to a 3.8% sodium citrate solution (9:1). The supernatant containing platelet-poor plasma was collected in conical tubes and stored at − 20 °C. Coagulation assays were performed on a semiautomatic coagulometer. To perform the tests, 100 µL of platelet-poor plasma was incubated at 37 °C for 5 min at different concentrations of xylan (6.25–200 µg/mL/20 µL) with 100 µL of APTT Pathromtin SL reagent. The clotting time and the anticoagulant activity was evaluated by detecting activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT). Sodium chloride 0.9% was used as a negative control, while heparin (6.25 µg/mL) was used as a positive control. All tests were performed in triplicate.

In vitro immunomodulatory activity

Cell viability assays in splenocyte cultures

Cytotoxicity assays were performed according to Santos et al. (2021) and Melo et al. (2022) with few modifications. Initially, splenocyte cultures (106 cells/well) were treated with different concentrations of xylans (6.25–200 μg/mL) for 24 h and incubated in a CO2 oven. Then, the cells were centrifuged, stained with propidium iodide (50 µM) and annexin V for 10 min and analyzed in flow cytometry (FACS Calibur platform) in 10,000 events. Data were analyzed using Flowing 2.0.1® software. Cell proliferation experiments were evaluated using CFSE under the same conditions as splenocyte cytotoxicity assays. The experimental control consisted only of cells and culture medium.

Investigation of cytokines and production of nitric oxide

The concentrations of cytokines produced in the supernatant of the cells stimulated with xylans were determined using the mouse Cytometric Array Kit (CBA) by flow cytometry. The nitric oxide released by the same cells was measured by the Griess method. The reading was performed in a microplate spectrophotometer (Multiskan FC, Thermo Scientific®) at 595 nm (Melo et al. 2022). The experimental control consisted of the supernatant of untreated cells.

Ethics Committee

All experimental procedures for hemolytic and anticoagulant activity assays and for obtaining splenocytes were carried out in accordance with the Ethics Committee on Animal Use of the Instituto Aggeu Magalhães/Fundação Oswaldo Cruz, protocol number 164/2020.

In vitro antioxidant activity

For the antioxidant activity tests, the xylans in water were subjected to a variation of 781–1000 µg. The DPPH assay was performed according to Zhang et al. (2019) and Boonchuay et al. (2021). For this, 0.1 mL of solutions were added to xylans in the DPPH solution (3.9 mL) and the system was incubated at 25 °C for 30 min and analyzed in a spectrophotometer at 515 nm. The blank of the equipment was ethanol. The ABTS assay was performed in the same proportions as the DPPH assay. However, the assays were incubated for 15 min and analyzed in a spectrophotometer at 734 nm.

The phosphomolybdenum complex reduction assay was performed according to Albuquerque Nery et al. (2022). For this, 3 mL of phosphomolybdenum complex, 6.9 mL of water and 0.1 mL of xylan solution were used. The blank was prepared by adding distilled water in place of the xylan solution. The tubes were incubated at 95 °C for 90 min and, after cooling, the reading was taken at 695 nm in a spectrophotometer. For the iron reduction assay, aliquots of 0.01 mL of xylan, at different concentrations, were diluted in 1 mL of distilled water and transferred to a tube containing 2.5 mL of 0.2 M phosphate buffer (pH 6.6) and 2.5 mL of 1% potassium ferricyanide (w/v). The mixture was incubated at 45 °C for 20 min. 2.5 mL aliquots of 10% (w/v) trichloroacetic acid were added. Then 2.5 mL aliquots of the mixture were added to 2.5 mL of distilled water and 0.5 mL of 0.1% (w/v) FeCl3. The absorbance reading was performed at 700 nm in a spectrophotometer. From an amount of antioxidant necessary to decrease the initial concentration by 50%, called IC50, it was determined in a non-linear way. All were performed in triplicate. The standards used were ascorbic acid and butylated hydroxytoluene (BHT).

Emulsifying activity (EA) and emulsion cream index (ECI)

The tests were carried out according to Wang et al. (2022) with few modifications. Initially the xylans were dissolved in water at 80 °C for 25 min at a concentration of 4%. Then, a serial dilution was performed, varying the xylan concentrations between 4.0 and 0.5%. The emulsions were prepared by mixing xylan and oil solutions in a ratio of 8:2, the system was homogenized for 10 min. In addition to evaluating the effect of xylan concentration, the influence of pH variation (3–11) and sodium ion concentration was also evaluated, using sodium chloride at different concentrations (0–200 µM) only at a concentration of 1% of xylans.

For the determination of EA, a volume of 100 μL of emulsion was added to 10 mL of a 0.1% aqueous solution of sodium dodecyl sulfate. Then, the absorbances were determined at 500 nm in a spectrophotometer. For the determination of the ECI, the emulsions were continuously centrifuged at 4000 rpm for 10 min until the emulsion was stratified, and the different centrifugation times were recorded. The ECI was calculated according to Eq. 3. The experimental control consisted of the system without xylan.

ECI=HAHT×100% 3

where HA is the height of the upper phase and HT is the total height of the emulsion.

In vitro prebiotic activity assays

The in vitro prebiotic activity of the xylans obtained in this study was evaluated by the methodology proposed by Thompson et al. (2022) with modifications. Two probiotic strains Lactobacillus casei ATCC 25,180 and Lactobacillus rhamnosus ATCC 7469 were cultivated in MRS broth and incubated at 37 °C for 24 h. After this period, the broth containing the bacteria was centrifuged at 11,000 rpm for 20 min at 4 °C. The cell precipitate was suspended in 0.85% saline and standardized to a cell concentration of 107 CFU/mL. Samples of xylans (PPB, PPL and commercial) and inulin were filtered through 0.22 μm filters and separately added to 100 mL of MRS broth to obtain a 0.25% solution. Therefore, four different tests were performed for Lactobacillus casei and four for Lactobacillus rhamnosus. The initial cell concentration was 106 CFU/mL and assays were incubated at 37 °C. Aliquots (1 mL) were withdrawn every 12 h for 48 h, diluted from 10−1 to 10−6 in saline solution (NaCL 0.85%). After dilution, a volume of 10 μL was inoculated in Petri plates containing MRS agar medium and incubated again at 35 °C for 24 h. Growth results were expressed in log CFU/mL. The experimental control used were systems containing inulin (a commercial prebiotic) and the standard used was commercial xylan.

Evaluation of the protective effect of xylans on the survival of probiotic yeasts Saccharomyces cerevisiae in simulation of gastrointestinal fluids

The gastric simulation tests were performed according to the methodology proposed by Larsen et al. (2018) with few modifications. The gastrointestinal simulation system was designed as follows: 20 mL of citrate/phosphate buffer, 4 mL of enzyme solution and 86 mL of gastric electrolyte solution. Saccharomyces cerevisiae were grown in Sabouraud culture medium for 24 h, then the system was centrifuged at 11,000 rpm for 10 min, the culture medium was discarded and the cells were resuspended in 0.85% NaCl solution to obtain a cell concentration of 1010 CFU/mL. Finally, 1 mL of cells was added to 9 mL of gastric solution and incubated at 37 °C under agitation (220 rpm) for 6 h. The incubation time used was adjusted to reflect the total period of food passage through the stomach and duodenum, simulating the digestive action of gastric and pancreatic fluids. Cell viability was determined by plating 10 µL of serial dilutions (10–1 to 10–6), in 0.85% NaCl solution, of the system in Petri dishes containing Sabouraud Agar and incubated at 35 °C for 48 h. The experiments were carried out in triplicate and as an experimental control (system without xylan), only the xylan-free gastric solution.

Statistical analysis

Results were expressed as mean ± standard error of the mean. Statistical analysis was performed using analysis of variance (ANOVA), with Tukey's test to assess significant differences between groups, p values < 0.05 were considered significant. All statistical analysis, were performed using the GraphPad 5.0 software.

Results and discussion

Analysis of the composition of branches and leaves of Protium puncticulatum

The composition analysis is related to the determination of cellulose, hemicellulose, lignin, extractives and ash contents. Figure 1 presents the composition in terms of percentage of these constituents for the branches and leaves of Protium puncticulatum.

Fig. 1.

Fig. 1

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Composition analysis results for branches (A) and leaves (B) of Protium puncticulatum respectively

The results presented in Fig. 1 show that the chemical composition between the branches and the leaves are different. The branches showed higher cellulose, lignin and ash contents when compared to the leaves that presented higher hemicellulose and extractive contents. According to Sluiter et al. (2010) and Ayeni et al. (2015), the chemical composition of plant material may vary according to seasonality, climatic conditions, study region, in addition to other factors. Even though there is a scarcity of studies on the composition of the species, Araújo (2019) reports the values of the composition of the wood of Protium puncticulatum, where he obtained the contents of cellulose 39%, lignin 27.88%, extractives 7.28% and ash 0.27%.

Yield of obtaining xylans

The yield results of obtaining xylans after lyophilization were 24.32 ± 0.1% for PPB xylans and 19.57 ± 0.3% for PPL xylans. These values are close to those obtained by Sharma et al. (2020a) that obtained 22.5% yield in xylan recovery from sawdust of Azadirachta indica (neem), Sharma et al. (2020b) that obtained yields of 23.5% for acacia sawdust xylan. Marques et al. (2019), optimizing the process of alkaline extraction of xylans from the bleached Kraft pulp of Eucalyptus globulus obtained yields ranging from 43 to 97%. Khaire et al. (2021) that obtained a 25% yield for sugarcane xylan.

Physicochemical characterization

Scanning electron microscopy (SEM)

Figure 2 shows the micrographs of the xylans evaluated in this study. The xylans PPB, PPL and commercial xylan showed similar morphology, i.e., a mixture of aggregated and non-aggregated particles with irregular morphology and rough surface. Similar results were obtained by Oliveira et al. (2010) evaluating corn cob xylans, Palaniappan et al. (2017) evaluating rice bran, millet and beech wood xylans and Khaire et al. (2021) for sugar cane xylan.

Fig. 2.

Fig. 2

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Micrographs of xylans PPB (A) and PPL (B) of and commercial (C), respectively, at 100× and 200× magnifications

Analysis of the composition of monosaccharides

The acid hydrolysis results showed that the evaluated xylans had only xylose units in their chemical composition, with values greater than 95% in their structure. Low levels of furfural were found in the analyzed hydrolysate, a product formed by the acid hydrolysis of C5 monosaccharides. Different works show that xylans have a high content of xylose in their structure. Khaire et al. (2021) found the presence of 74% of xylose residues, 16% (w/w) of glucuronic acid residues and 10% (w/w) of arabinose for sugar cane xylan. Lu et al. (2022) characterizing a commercial xylan from Betula pendula obtained the following composition: xylose (79.3%), 4-O-Me-GlcA (9.3%), galactose (3.7%), rhamnose (2.4%), mannose (2.0%), glucose (1.8%), Arabinose (1.8%), galacturonic acid (1.8%) and glucuronic acid (0.5%).

Elementary analysis

The elemental percentage composition of PPB, PPL and commercial xylans were similar in terms of C and H. For carbon, the following values were obtained: 34.94 ± 0.3, 33.79 ± 0.1 and 34.79 ± 0.1% and 6.34 ± 0.02, 5.79 ± 0.01 and 5.47 ± 0.2% for hydrogen respectively. Nitrogen or sulfur contents were found in none of the samples, which indicate the purity of the isolated xylans (Sharma et al. 2020a, b). The percentage values obtained for oxygen were: 58.72 ± 0.1; 60.42 ± 0.3 and 59.74 ± 0.5 for xylans PPB, PPL and commercial respectively.

These results were close to those obtained by Palaniappan et al. (2017) evaluating the elemental percentage composition of xylans extracted in water obtained from rice bran and millet tegument compared to wood xylans, obtained percentages of 34.83, 33.55 and 34.66 for carbon and 3.51, 4.57 and 5.44 for hydrogen. In addition, values of N (1.01, 2.76 and 1.03) greater than S (0.12, 0.47 and 0.14) were obtained, which were attributed to traces of proteins or phenolic compounds present in the structure.

Sharma et al. (2020b) performing the analysis of the elemental composition of the xylan extracted from acacia sawdust found that the polysaccharide contained 36.70% carbon and 3.92% hydrogen, while the nitrogen and sulfur contents are absent in the extracted xylan, confirming that the extracted xylan is highly pure.

Analysis by total reflection attenuated infrared spectroscopy with Fourier transform (ATR-FTIR) and X-ray diffraction (XRD)

Figure 3A shows the ATR-FTIR. The bands were previously identified by Sharma et al. (2020a), Sharma et al. (2020b) and Khaire et al. (2021), presenting different xylans. Figure 3B shows the diffractograms for PPB, PPL and commercial xylan, respectively.

Fig. 3.

Fig. 3

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ATR-FTIR spectrum (A) and diffractograms (B) for PPB PPL and commercial xylans respectively

Figure 3A shows the bands at 3700 − 3000 cm−1 correspond to the vibrations of the OH groups involved in hydrogen bonds. Symmetric stretching of CH in xylan was observed by the band in the region from 2936 to 2920 cm−1. The bands at 1420 and 1310 cm−1 were associated with C–H, OH, or CH2 bending vibrations. The band at 1620 cm−1 refers to the bending of the O–H bond referring to the absorption of the molecule in water, in the case of water-soluble xylans. The bands between 1175 and 1000 cm−1 refer to the stretching of the C–O, C–C, C–OH and CO–C bonds. The bands with absorption at 897 − 890 cm−1, referring to the stretching and deformation of the C–O–C, C–C-O and C–CH bonds in the case of xylans, the β-glycosidic bonds between the xylanopyranoses of the main chain.

The results presented in Fig. 3B show that the xylans showed a broad peak in the range of 20–22°, indicating that the structure of the evaluated polysaccharides is amorphous, in addition, no clear crystalline peak was observed. This profile was also observed by Sheikhi et al. (2018) evaluating a xylan isolated from sugarcane bagasse by two conventional alkaline extraction methods, using NaOH and KOH. Palaniappan et al. (2017) studied X-ray diffraction (XRD) patterns for xylans obtained from rice bran, millet seed coating and beechwood, and observed that these had a wider peak at 20° suggesting that xylan structures evaluated were completely amorphous. Fröhlich et al. (2022), evaluating quaternized (QXy) and sulfated (SXy) xylans produced from corn cob xylan, observed that the xylans obtained showed signs in a region between 18 and 20º.

Analysis by nuclear magnetic resonance spectroscopy (1H NMR,13CNMR and 1H-.13C HSQC)

Figure 4 shows the 1H NMR and 13 C NMR spectra for PPB, PPL and commercial xylans. The signs were previously identified by Lahaye et al. (2003), Bhanja et al. (2022), Wang et al. (2022) and Puițel et al. (2022), evaluating different xylans.

Fig. 4.

Fig. 4

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1H NMR (A) and 13C NMR (B) spectrum for PPB PPL and commercial xylans, respectively

1H NMR (Fig. 4A) spectra showed signals in the region of 3.0–4.4 ppm attributed to d-xylose. signs were also observed at 4.34; 3.97; 3.62; 3.51; 3.25 and 3.04 ppm referring to H-1, H-5equatorial, H-4, H-3, H-5axial and H-2 present in the xylose structure. Signals in the region at 4.3–5.9 ppm are attributed to the chemical shift of the anomeric H. In this region it was possible to observe the presence of signals attributed to α-anomeric H (4.9–5.4 ppm) and to β-anomeric H (4.3–4.9 ppm). The spectra were shown in Fig. 4B and help to elucidate the carbonic structure of these polysaccharides. The spectra showed characteristic signals C-1, 2, 3, 4 and 5; δ = 102.5; 73.5; 74.5; 77.2 and 63.8 ppm. Furthermore, the spectra of PPB and PPL xylans showed structural similarities.

After 1D NMR analysis the xylans were characterized by 2D HSQC NMR for further investigation. Figure 5 shows the 2D HSQC NMR spectra for PPB (Fig. 5A), PPL (Fig. 5B) and commercial xylans (Fig. 5C) respectively. The signs were previously identified by Zhang et al. (2022), Bhanja et al. (2022) and Liu et al. (2021) respectively.

Fig. 5.

Fig. 5

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HSQC NMR spectrum for PPB (A), PPL (B) and commercial xylans (C), respectively

The HSQC spectra of xylans PPB (Fig. 5A) and PPL (Fig. 5B) were similar. Five predominant cross-signals of units linked to β-d-Xyl p-(1 → 4) were determined and these were detected in 101.64/4.42; 72.67/3.23; 73.66/3.49; 76.33/3.72 and 62.93/3.32 ppm, which are characteristic of C1 single bond H1, C2 single bond H2, C3 single bond H3, C4 single bond H4 and C5 single bond H5, respectively. In contrast, the signals obtained for commercial xylan (Fig. 5C) were 72.67/3.23 (C2 single bond H2), 76.33/3.72 (C4 single bond H4) and 62.93/3.32 (C5 single bond H5) respectively.

Thermogravimetric and differential scanning calorimetry (DSC) analysis

Figure 6 shows the mass loss curves thermogravimetric analysis (TGA) (Fig. 6A) the mass loss rate represented by the first derivative (DTG) (Fig. 6B), and the differential scanning calorimetry (DSC) curves (Fig. 6C) obtained for the xylans in this study.

Fig. 6.

Fig. 6

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Thermogravimetric (A) and thermogravimetric derivative (B) analysis curves for the xylans PPB, PPL and commercial

The curves presented in Fig. 6A and 6B showed that the xylans obtained in this study and the commercial xylan presented three stages of thermal degradation. The first decomposition stage occurred in the range of 35 to 160 °C, and corresponds to the loss of moisture content or water content. The second stage at 170–350 °C takes place the thermal decomposition of xylan itself into smaller molecules such as CH3COOH, CH4, CO, CO2 and HCOOH. Finally, the third stage at temperatures higher than 350 °C is the degradation of residual xylan remaining in ash. Similar results were obtained by Bahcegul et al. (2013), Jin et al. (2019) and Khaire et al. (2021) evaluating the thermal behavior of different xylans.

The curves presented in Fig. 6C (DSC) for the evaluated xylans showed similar profiles evidencing an endothermic peak in the temperature range of 50–110 °C, indicating the loss of water from the xylan and an exothermic peak in the range of 250 to 280 °C related to thermal decomposition. The enthalpy values at the highest temperature of degradation (279 ºC) were 110.19 J/g, 108.01 J/g and 126.60 J/g for xylan PPB, PPL and commercial respectively. Similar results to the xylans evaluated in our study were obtained by Ünlü et al. (2015), Costa Urtiga et al. (2020), Seera et al. (2021) evaluating different xylans.

Molecular mass of xylans

The number average (Mn), weight (Mw) and polydispersity (Mw/Mn) were obtained by gel permeation chromatography. The Mn, Mw and Mw/Mn results for the xylan PPB were 25.10 g/mol; 35.89 g/mol and 1.42 for PPL 24.89 g/mol; 35.74 g/mol and 1.4 and for commercial xylan it was 23.56 g/mol; 37.84 g/mol and 1.6 respectively. The close polydispersion indices indicate that these xylans have similar molecular mass fragments.

The literature presents different values for the molecular weight of xylans. Liu et al. (2021) evaluating xylans obtained from corn stalk granules, corn stalk strips and corn stalk filaments, obtained Mn values ranging from 72.054 to 74.033 g/mol, Mw ranging from 28.565 to 32.890 g/mol and Mw/Mn ranging from 2.22 to 2.52.

Cytotoxicity against J774 macrophage cells, V79 cells, Vero cells and erythrocytes

Figure 7 shows the toxicity profile of xylans PPB, PPL and commercial xylan against J774 macrophage cells (Fig. 7A), V79 cells (Fig. 7B), Vero cells (Fig. 7C) and erythrocytes (Fig. 7D).

Fig. 7.

Fig. 7

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Results of cytotoxicity promoted by xylans PPB, PPL and commercial, against macrophage cells J774 (A), V79 cells (B), Vero cells (C) and erythrocytes (D) respectively

The results presented show that the evaluated xylans were not cytotoxic against J774 macrophage cells (Fig. 7A), V79 cells (Fig. 7B), Vero cells (Fig. 7C) showing cell viability greater than 95%. Furthermore, the evaluated xylans showed hemolysis values lower than 10%, indicating that these polysaccharides are not hemolytic either (Fig. 7D).

Similar results were obtained by Sharma et al. (2020a) who found that xylan from sawdust from Azadirachta indica (neem) had no cytotoxic effect against normal fibroblast cells at concentrations ranging from 0.05–0.2 mg/mL. Melo-Silveira et al. (2019) verified that corn cob xylans did not promote cytotoxic effects against murine macrophage cells (RAW 264.7) at concentrations ranging from 0 to 2.0 mg/mL. Mandal et al. (2010) verified that Scinaia hatei xylans did not promote a cytotoxic effect against Vero cells. Finally, Kumar et al. (2021) found that xylan-based nanoparticles promoted a low percentage of hemolysis at concentrations ranging from 0 to 2.0 mg/mL.

Anticoagulant activity in vitro

Anticoagulant activity was evaluated by classical coagulation assays for APTT, PT and TT these parameters play different roles in the analysis of hemostasis (Chen et al. 2020). PT is an important index to demonstrate the abnormality of extrinsic blood clotting. The APTT can be used to verify the intrinsic blood clotting pathway, and the TT is used to assess whether there is a certain anticoagulant property in the blood when PT and APTT are prolonged (Melo-Silveira et al. 2011).

Table 1 (in supplementary material) presents the results of APTT, TT and PT for the xylans PPB, from PPL and for the commercial xylan, respectively. The results presented in Table 1 show that the xylans evaluated here were able to promote an increase in the time of coagulation parameters with increasing concentration in a similar way. However, even with this increase, xylans showed shorter time results when compared to standard heparin. The results presented for the xylans evaluated in this study show that the differences were not significant.

Similar results were obtained by Melo-Silveira et al. (2011) evaluating corn cob xylans. Liang et al. (2015) obtained promising results for β-1,3-xylan extracted from Caulerpa lentillifera, the authors found that the evaluated polysaccharide was able to promote an increase in clotting time. In addition to in natura xylans, different studies have been proposed with modified xylans to promote increased anticoagulant activity. Cai et al. (2015) and Chen et al. (2020) found that sulfated sugarcane bagasse xylans were able to promote an increase in clotting time. Fras Zemljič et al. (2020) evaluating sulfated 4-O-methyl glucuronoxylan from beech wood also found an increase in clotting time.

Evaluation of in vitro immunomodulatory activity

Figure 8 shows the cytotoxicity values promoted by the xylans under study using the annexin V (Fig. 8A) and propidium iodide (Fig. 8B) methods to assess cell death.

Fig. 8.

Fig. 8

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Viability results promoted by xylans PPB, PPL and commercial xylan against splenocyte cells evaluated by staining with annexin V (A) and propidium iodide (B)

Spleen cells from Balb/c mice treated with xylans at different concentrations did not undergo significant cell death at any of the concentrations evaluated. In fact, xylans PPB and PPL and commercial xylan promoted high cell viability with values greater than 95%. Similar results were obtained by Shi et al. (2014) evaluating the cytotoxic effect of a xylan obtained from the roots of Cudrania tricuspidata. After evaluating the cytotoxicity, the proliferative effect promoted by the xylans under study was evaluated, through the method using CFSE. This method is based on the geometric progression of CFSE fluorochrome decay in cells. Assays were performed at concentrations ranging from 6.25 to 200 µg/mL. Figure 9 shows the results of the proliferative effect promoted by the evaluated xylans.

Fig. 9.

Fig. 9

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cell proliferation results using CFSE, xylans PPB, PPL and commercial xylan

The results showed that splenocytes were able to proliferate when in contact with the xylans evaluated in this study. Similar results were obtained by Shi et al. (2014) and Bhanja et al. (2022) evaluating the effect of xylans at different concentrations. In addition to the proliferative assays, assays were performed for the determination of cytokines and nitric oxide (in the cell culture supernatant). For the determination of cytokines, only a concentration of 12.5 µg/mL was used, a concentration previously used for the determination of cytotoxins produced by different polysaccharides as reported by Santos et al. (2021) and Melo et al. (2022). Furthermore, this is a non-toxic concentration for splenocytes and not for animal cells evaluated in this study. The results of the production of cytokines and nitric oxide were presented in Table 2 (in supplementary material).

The results presented in Table 2 show that xylans were able to induce an increase in cytokine production. Thus, it was possible to observe the profile of the predominant pro-inflammatory response, through the higher levels of cytokines characteristic of the pro-inflammatory response (IL-2, and TNF-α and IFN-γ), when compared to the profile of the cytokine profile. anti-inflammatory (IL-4). In addition, a non-significant increase in nitric oxide was observed. Data found in the literature reveal that the chemical structure of macromolecules can be directly linked to the immunological activity (Ebringerová et al. 2002). Mendis et al. (2017) observed that an arabinoxylan polysaccharide increased the production of pro-inflammatory cytokines, IL-8 and TNF-α. Pynam et al. (2019) verified that a xylorhamnoarabinogalactan I was able to induce the immunoregulatory cytokines IL10/IL17.

In vitro anti-tumor activity

Figure 10 shows the results of antiproliferative activity promoted by xylans PPB, PPL and commercial xylan in Jurkat cells, MCF-7, DU-145, T47D and HepG2 respectively.

Fig. 10.

Fig. 10

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Results of antitumor activity promoted by xylans PPB, PPL and commercial against cells against different cancer cells Jurkat (A), MCF-7 (B), DU-145 (C), T47D (D) and HepG2 (E)

The results presented in Fig. 10 showed that xylans were able to promote increased cytotoxicity with increasing concentration. Similar behavior was obtained by Melo-Silveira et al. (2011), Melo-Silveira et al. (2019) and Li et al. (2021) evaluating different xylans against different tumor cells. It was possible to observe low activity results, that is, inhibition values lower than 50%, not being able to determine the IC50.

The literature reports xylans as potential antiproliferative agents against different tumor cell lines. Melo-Silveira et al. (2019) evaluating the antiproliferative activity of a corn cob xylan found that it was able to inhibit the proliferation of human lung adenocarcinoma epithelial cells (A549) by 20% and human cervical adenocarcinoma cells (HeLa) by 60% at a concentration of 2 mg/mL (2000 µg/mL).

Qian et al. (2022) found that the antitumor activity is directly related to the chemical structure of xylans, evaluated the inhibitory effects of corn straw xylans (XYCS) in natura and phosphorylated (XYCS-P) against HepG2 cells, in concentrations ranging from 50 to 400 µg/mL. The authors found that cell viability decreased from 72.42 to 47.7%, while the cell viability of XYCS at 400 μg/mL was 64.52%, demonstrating that XYCS-P significantly improved the inhibitory effects for HepG2 cells. IC50 and XYCS-P were determined to be 335.91 μg/mL.

In vitro antioxidant activity

Xylans are polysaccharides capable of promoting antioxidant activity in vitro (Melo-Silveira et al. 2011; Zhang et al. 2019; Boonchuay et al. 2021). Although the mechanism of its antioxidant action is not yet fully established, it is known that the hydroxyl groups present in the structure of the molecule can present themselves as hydrogen donors (Zhang et al. 2021). In addition to these groups, other characteristics such as molecular weight and levels of phenolic compounds (obtained during the extraction process) may contribute to the activity (Renault et al. 2014; Zhang et al. 2021).

Figure 11 shows the in vitro antioxidant activity curves in different assays promoted by xylans PPB and PPL compared to commercial xylan and to ascorbic acid and BHT standards at different concentrations.

Fig. 11.

Fig. 11

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Antioxidant activity promoted by xylans PPB, PPL compared to commercial xylan and to ascorbic acid and BHT standards by different in vitro methods DPPH (A), ABTS (B), reduction of the phosphomolybdenum complex (C) and reduction of ions iron (D) respectively

The curves obtained in Fig. 11 show that the xylans PPB, PPL and commercial xylan promoted low results of antioxidant activity in vitro. For the DPPH radical capture assay (Fig. 11A) xylans were able to promote activity results that varied around 10%. The ABTS radical capture assay (Fig. 11B) showed better results when compared to the DPPH results, with a percentage of capture ranging from 30 to 35.9%.

The xylans were able to reduce the phosphomolybdenum complex (Fig. 11C) with values lower than 6%. Finally, xylans were able to reduce iron ions (Fig. 11D). In addition to these low values of antioxidant activity, the xylans did not present phenolic constituents in their structure. These results indicate that antioxidant activity was promoted only by xylans, and indicate the purity of the molecules. Silva (2019) obtaining sugarcane bagasse xylans found that the evaluated polysaccharides were not able to promote antioxidant activity, this may be related to the lower concentration of phenolic compounds present in the sample.

Emulsifying activity

Figure 12 presents the results of emulsifying activity (EA) and emulsion cream index (ECI) under different experimental conditions.

Fig. 12.

Fig. 12

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Results of emulsifying activity and cream index promoted by xylans PPB, PPLand commercial. Concentration variation (A and D). At a concentration of 1%, variation in the concentration of NaOH (B and E) and pH (C and F) respectively

The results showed that the evaluated xylans were able to promote emulsification at different concentrations (Fig. 12A). Furthermore, it was observed that the concentration of 2% presented similar results to those found in the concentration of 4%, so the concentration of 2% was chosen for the verification tests the influence of the concentration of sodium ions and the pH. The results presented in Fig. 12B showed that the concentration of sodium chloride did not significantly influence the emulsion. Regarding the pH (Fig. 12C) it was verified that the emulsion values were close, highlighting those obtained at pH 7.0. Regarding the emulsion cream index, all the results presented in Fig. 12D–F were low.

The literature presents different results for the emulsifying activity promoted by xylans. The emulsification mechanism promoted by xylans is complex and this is directly related to the chemical structure of this polysaccharide (Tang et al. 2021; Wang et al. 2022). The structure of these molecules has many hydrophilic groups, such as hydroxyl and carboxyl, which confers greater solubility (giving the structure a more polar character). However, in addition to the hydrophilic groups, these polysaccharides present (to a lesser extent) hydrophobic groups which are directly related to the emulsifying activity (Tang et al. 2021).

The emulsifying potential of xylans was also observed by Wang et al. (2022) evaluating different natural and acetylated xylans. The authors found that the evaluated polysaccharides showed promising results in terms of emulsifying activity and emulsion cream index. Hu et al. (2021) when functionalizing a glucuronoxylan with amino groups, found that these polysaccharides were able to promote an increase in emulsifier activity.

Evaluation of prebiotic activity in vitro

Figure 13 shows the results of growth in individual cultures of Lactobacillus in culture media with the carbon source of inulin (commercial prebiotic) xylans PPB, PPL and commercial in a growth ranging from 0 to 48 h.

Fig. 13.

Fig. 13.

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Evaluation of prebiotic activity in vitro. Growth of L. paracasei (A) and L. rhamnosus (B) on inulin, xylans PPB, PPL and commercial

The results presented in Fig. 13 showed that the xylans PPB, PPL and commercial were able to promote the growth of the microorganisms L. paracasei (Fig. 13A) and L. rhamnosus (Fig. 13B) in a period of 48 h in a similar way. The prebiotic potential of xylans is associated with the mechanism (Zhang et al. 2022). During microbial growth, xylans are hydrolyzed by different xylanases that, during metabolism, are degraded into short-chain fatty acids (acetic, lactic butyric and propionic acids) and different gases. This microenvironment promotes beneficial effects on human and animal health (Singh et al. 2015; Zhang et al. 2022). Similar results were obtained by Yan et al. (2018) evaluating different xylooligosaccharides obtained from industrial xylan residue, found that the evaluated polysaccharides were able to promote the growth of microorganisms Bifidobacteria adolescents and Lactobacillus acidophilus.

Ravichandra et al. (2022) evaluating xylooligosaccharides derived from sorghum xylan from sugarcane, the authors found that the evaluated polysaccharides promoted the growth of Lactobacillus T-10. Ríos-Ríos et al. (2022) verified that xylo-oligosaccharides obtained from beech wood xylan promoted the growth of Lactobacillus brevis DSM 20054, Bifidobacterium adolescentis DSM20083 and Escherichia coli K88 respectively.

Evaluation of the protective effect of xylans on the survival of Saccharomyces cerevisiae yeasts in gastrointestinal conditions

Figure 14 shows the cell viability results in the gastrointestinal simulation assay in the presence or absence of xylans in a period of 6 h.

Fig. 14.

Fig. 14.

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Gastric simulation in a system containing yeasts and PPB, PPL and commercial xylans

The results showed that cell viability decreased during the 6 h in contact with the gastrointestinal solution, with a reduction > 2.0 Logs. However, it was possible to observe that xylan-containing systems promoted the reduction of viability loss to < 2.0 Logs, making the yeast + xylan system resistant to gastrointestinal conditions (Alkalbani et al. 2019). This increase in viability may be associated with specific interactions of xylans with different structures present on the surface of the microorganism (Larsen et al. 2018). As the chemical structures of the evaluated xylans show similarities, the results showed no significant difference. Similar results were observed by Wu et al. (2018), Yasmin et al. (2019) and Ravichandra et al. (2022), evaluating different polysaccharides.

Conclusion

The xylans evaluated in this study can be classified as homoxylans. These polysaccharides showed low cytotoxicity against animal cells and were able to inhibit the growth of tumor cell lines. As for immunological assays, xylans were able to stimulate the growth of immune cells in addition to inducing the production of pro-inflammatory cytokines, being considered immunomodulatory agents. Antioxidant activity assays showed that xylans have low activity. Furthermore, the polysaccharides in this study were able to promote emulsification in xylan–oil systems. In prebiotic activity assays, it was observed that these polysaccharides may be promising prebiotic agents promoting the growth of different Lactobacillus. Finally, it was observed that these polysaccharides were able to protect probiotic yeasts of the species Saccharomyces cerevisiae against gastrointestinal conditions. Therefore, it is possible to conclude that the xylans obtained from the branches and leaves of Protium puncticulatum are promising polysaccharides to be used as raw material for the creation of products applied for biomedical and food purposes, due to the different biological properties capable of promoting benefits to health.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 18 kb) (18.1KB, docx)

Acknowledgements

This study was funded by Brazilian agencies FACEPE (Process APQ-0498-4.03/19), researcher retention grant FACEPE (Process BFP-0038-04.03/21) and CNPq (Process 306865/2020-3). In addition to these, we would also like to thank Mil Madeiras Preciosas, for providing samples of the branches and leaves of Protium puncticulatum.

Data availability

The authors made available the data presented in this work.

Declarations

Conflict of interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References

  1. Albuquerque Nerys LL, Jacob ITT, Silva AR, Oliveira AM, Rocha WRV, Pereira DTM, Abreu AS, Silva RSM, Cruz Filho IJC, Lima MDCA. Photoprotective, biological activities and chemical composition of the non-toxic hydroalcoholic extract of Clarisia racemosa with cosmetic and pharmaceutical applications. Ind Crops Prod. 2022;180:114762. doi: 10.1016/j.indcrop.2022. [DOI] [Google Scholar]
  2. Alkalbani NS, Turner MS, Ayyash MM. Isolation, identification, and potential probiotic characterization of isolated lactic acid bacteria and in vitro investigation of the cytotoxicity, antioxidant, and antidiabetic activities in fermented sausage. Microb Cell Fact. 2019;18(1):1–12. doi: 10.1186/s12934-019-1239-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Araújo RD (2019) Avaliação do potencial de espécies manejadas na Amazônia para produção de painéis “EGP” (Edge Glued Panels) não estrutural.
  4. Arruda MDM, Alves SDPL, Cruz Filho IJ, Sousa GF, Souza Silva GA, Nascimento Santos DKD, Melo CML. Characterization of a lignin from Crataeva tapia leaves and potential applications in medicinal and cosmetic formulations. Int J Biol Macromol. 2021;180:286–298. doi: 10.1016/j.ijbiomac.2021.03.077. [DOI] [PubMed] [Google Scholar]
  5. Ayeni AO, Adeeyo OA, Oresegun OM, Oladimeji TE. Compositional analysis of lignocellulosic materials: Evaluation of an economically viable method suitable for woody and non-woody biomass. Am J Eng Technol Res. 2015;4(4):14–19. doi: 10.1016/j.ijbiomac.2021.03.077. [DOI] [Google Scholar]
  6. Bahcegul E, Akinalan B, Toraman HE, Erdemir D, Ozkan N, Bakir U. Extrusion of xylans extracted from corn cobs into biodegradable polymeric materials. Bioresour Technol. 2013;149:582–585. doi: 10.1016/j.biortech.2013.09.097. [DOI] [PubMed] [Google Scholar]
  7. Bhanja SK, Maity P, Rout D, Sen IK, Patra S. A xylan from fresh Piper betle leaves: structural characterization and studies of bioactive properties. Carbohydr Polym. 2022;291:119570. doi: 10.1016/j.carbpol.2022.119570. [DOI] [PubMed] [Google Scholar]
  8. Boonchuay P, Wongpoomchai R, Jaturasitha S, Mahatheeranont S, Watanabe M, Chaiyaso T. Prebiotic properties, antioxidant activity, and acute oral toxicity of xylooligosaccharides derived enzymatically from corncob. Food Biosci. 2021;40:100895. doi: 10.1016/j.fbio.2021.100895. [DOI] [Google Scholar]
  9. Cai W, Chen Q, Xie L, Yang L, Zhang R. Extraction, sulfonation and anticoagulant activity of xylan from corncob. Eur Food Res Technol. 2015;240(5):969–975. doi: 10.1007/s00217-014-2401-y. [DOI] [Google Scholar]
  10. Chen Z, Li S, Fu Y, Li C, Chen D, Chen H. Arabinoxylan structural characteristics, interaction with gut microbiota and potential health functions. J Funct Foods. 2019;54:536–555. doi: 10.1016/j.jff.2019.02.007. [DOI] [Google Scholar]
  11. Chen Y, Sun X, Shan J, Tang C, Hu R, Shen T, Ying H. Flow synthesis, characterization, anticoagulant activity of xylan sulfate from sugarcane bagasse. Int J Biol Macromol. 2020;155:1460–1467. doi: 10.1016/j.ijbiomac.2019.11.124. [DOI] [PubMed] [Google Scholar]
  12. Costa Urtiga SC, Alves VMO, Oliveira Melo C, Lima MN, Souza E, Cunha AP, Egito EST. Xylan microparticles for controlled release of mesalamine: production and physicochemical characterization. Carbohydr Polym. 2020;250:116929. doi: 10.1016/j.carbpol.2020.116929. [DOI] [PubMed] [Google Scholar]
  13. Deutschmann R, Dekker RF. From plant biomass to bio-based chemicals: latest developments in xylan research. Biotechnol Adv. 2012;30(6):1627–1640. doi: 10.1016/j.biotechadv.2012.07.001. [DOI] [PubMed] [Google Scholar]
  14. Ebringerová A, Kardošová A, Hromádková Z, Malovíková A, Hříbalová V. Immunomodulatory activity of acidic xylans in relation to their structural and molecular properties. Int J Biol Macromol. 2002;30(1):1–6. doi: 10.1016/S0141-8130(01)00186-6. [DOI] [PubMed] [Google Scholar]
  15. Ebringerová A, Hromádková Z, Košťálová Z, Sasinková V. Chemical valorization of agricultural by-products: isolation and characterization of xylan-based antioxidants from almond shell biomass. Resources. 2008;3(1):60–70. [Google Scholar]
  16. Fras Zemljič L, Dimitrušev N, Zaplotnik R, Strnad S. Insights into adsorption characterization of sulfated xylans onto poly (ethylene terephthalate) Polymers. 2020;12(4):825. doi: 10.3390/polym12040825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hu Z, Wang C, Xiang Z, Lu F. Amino-functionalized glucuronoxylan as an efficient bio-based emulsifier. Cellulose. 2021;28(6):3677–3689. doi: 10.1007/s10570-021-03762-5. [DOI] [Google Scholar]
  18. Iwakiri S, Trianoski R, Fonte APN, França MC, Lau PC, Molleken R (2016) Potential use of the wood of Dinizia excelsa Ducke and Protium puncticulatum J. F. Macbr for manufacture of EGP panels. Sci For 111: 709–7017. http://www.ipef.br/publicacoes/scientia/.
  19. Jana UK, Kango N, Pletschke B. Hemicellulose-derived oligosaccharides: emerging prebiotics in disease alleviation. Nutr Front. 2021 doi: 10.3389/fnut.2021.670817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jin X, Hu Z, Wu S, Song T, Yue F, Xiang Z. Promoting the material properties of xylan-type hemicelluloses from the extraction step. Carbohydr Polym. 2019;215:235–245. doi: 10.1016/j.carbpol.2019.03.092. [DOI] [PubMed] [Google Scholar]
  21. Khaire KC, Sharma K, Thakur A, Moholkar VS, Goyal A. Extraction and characterization of xylan from sugarcane tops as a potential commercial substrate. JBB. 2021;131(6):647–654. doi: 10.1016/j.jbiosc.2021.01.009. [DOI] [PubMed] [Google Scholar]
  22. Kumar A, Kumar B, Kulshreshtha A, Negi YS. Redox-sensitive nanoparticles based on xylan-lipoic acid conjugate for tumor targeted drug delivery of niclosamide in cancer therapy. Carbohydr. Res. 2021 doi: 10.1016/j.carres.2020.108222. [DOI] [PubMed] [Google Scholar]
  23. Lahaye M, Rondeau-Mouro C, Deniaud E, Buléon A. Solid-state 13C NMR spectroscopy studies of xylans in the cell wall of Palmaria palmata (L. Kuntze, Rhodophyta) Carbohydr. Res. 2003;338(15):1559–1569. doi: 10.1016/S0008-6215(03)00241-6. [DOI] [PubMed] [Google Scholar]
  24. Li M, Li H, Liu H, Zou Z, Xie C. Synthesis and biological evaluation of a novel glycidyl metharcylate/phaytic acid-based on bagasse xylan composite derivative. Polymers. 2021;13(13):2084. doi: 10.3390/polym13132084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Liang WS, Liu TC, Chang CJ, Pan CL (2015) Bioactivity of β-1, 3-xylan extracted from Caulerpa lentillifera by using Escherichia coli ClearColi BL21 (DE3)-β-1, 3-xylanase XYLII. J Food Nutr Res 3:437–444. 10.12691/jfnr-3-7-5
  26. Liu Q, Fan H, Mou H, Liu J, Huang J, Dong X, Song H. Preparation and characterization of xylan by an efficient approach with mechanical pretreatments. Ind Crops Prod. 2021;165:113420. doi: 10.1016/j.indcrop.2021.113420. [DOI] [Google Scholar]
  27. Mandal P, Pujol CA, Damonte EB, Ghosh T, Ray B. Xylans from Scinaia hatei: structural features, sulfation and anti-HSV activity. Int J Biol Macromol. 2010;46(2):173–178. doi: 10.1016/j.ijbiomac.2009.12.003. [DOI] [PubMed] [Google Scholar]
  28. Marques AI, Serrano MDL, Alves AMB, Sousa APM. Isolation of xylans from bleached Eucalyptus kraft pulp by antisolvents precipitation. Cellulose. 2019;26(3):1977–1992. doi: 10.1007/s10570-018-1941-x. [DOI] [Google Scholar]
  29. Melo CMLD, Sousa GFD, Silva GADS, Silva RSD, Bezerra Júnior NDS, Santos DKDDN, Lima MCA, Cruz Filho IJD. Pectin-like polysaccharide extracted from the leaves Caesalpinia pulcherrima is a promising antioxidant and immunomodulator agent. Braz Arch Biol Technol. 2022 doi: 10.1590/1678-4324-2022200718. [DOI] [Google Scholar]
  30. Melo-Silveira RF, Fidelis GP, Costa MSSP, Telles CBS, Dantas-Santos N, Oliveira Elias S, Ribeiro VB, Barth AL, Macedo AJ, Leite EL, Rocha HAO. In vitro antioxidant, anticoagulant and antimicrobial activity and in inhibition of cancer cell proliferation by xylan extracted from corn cobs. Int J Mol Sci. 2011;13(1):409–426. doi: 10.3390/ijms13010409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Melo-Silveira RF, Viana RLS, Sabry DA, Silva RA, Machado D, Nascimento AKL, Scortecci KC, Ferreira-Halder CV, Sassaki GL, Rocha HAO. Antiproliferative xylan from corn cobs induces apoptosis in tumor cells. Carbohydr Polym. 2019;210:245–253. doi: 10.1016/j.carbpol.2019.01.073. [DOI] [PubMed] [Google Scholar]
  32. Mendis M, Leclerc E, Simsek S. Arabinoxylan hydrolyzates as immunomodulators in Caco-2 and HT-29 colon cancer cell lines. Food Funct. 2017;8(1):220–231. doi: 10.1039/C6FO00866F. [DOI] [PubMed] [Google Scholar]
  33. Naidu DS, Hlangothi SP, John MJ. Bio-based products from xylan: a review. Carbohydr Polym. 2018;179:28–41. doi: 10.1016/j.carbpol.2017.09.064. [DOI] [PubMed] [Google Scholar]
  34. Palaniappan A, Yuvaraj SS, Sonaimuthu S, Antony U. Characterization of xylan from rice bran and finger millet seed coat for functional food applications. J. Cereal Sci. 2017;75:296–305. doi: 10.1016/j.jcs.2017.03.032. [DOI] [Google Scholar]
  35. Puițel AC, Suditu GD, Danu M, Ailiesei GL, Nechita MT. An experimental study on the hot alkali extraction of xylan-based hemicelluloses from wheat straw and corn stalks and optimization methods. Polymers. 2022;14(9):1662. doi: 10.3390/polym14091662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pynam H, Dharmesh SM. A xylorhamnoarabinogalactan I from Bael (Aegle marmelos L.) modulates UV/DMBA induced skin cancer via galectin-3 and gut microbiota. J Funct Foods. 2019;60:103425. doi: 10.1016/j.jff.2019.103425. [DOI] [Google Scholar]
  37. Qian S, Yu Y, Ma J, Diao E, Ye S, Gao J, Liu Y, Hu W. Evaluation of a novel phosphorylated corn straw xylan for enhancement of thermal stability, crystallinity and functional activity. Int J Food Sci. 2022;57(1):481–488. doi: 10.1111/ijfs.15428. [DOI] [Google Scholar]
  38. Ravichandra K, Balaji R, Devarapalli K, Batchu UR, Thadikamala S, Banoth L, Pinnamaneni SR, Prakasham RS. Enzymatic production of prebiotic xylooligosaccharides from sorghum (Sorghum bicolor (L.) xylan: value addition to sorghum bagasse. Biorefinery Biomass Convers. 2022 doi: 10.1007/s13399-021-02216-z. [DOI] [Google Scholar]
  39. Renault E, Barbat-Rogeon A, Chaleix V, Calliste CA, Colas C, Gloaguen V. Partial structural characterization and antioxidant activity of a phenolic–xylan from Castanea sativa hardwood. Int J Biol Macromol. 2014;70:373–380. doi: 10.1016/j.ijbiomac.2014.07.005. [DOI] [PubMed] [Google Scholar]
  40. Santos DKD, Silva Barros BR, Cruz Filho IJ, Júnior NDSB, Silva PR, Bomfim Nascimento PH, Lima MCA, Napoleão TH, Melo CML. Pectin-like polysaccharide extracted from the leaves of Conocarpus erectus Linnaeus promotes antioxidant, immunomodulatory and prebiotic effects. Bioact. 2021;26:100263. doi: 10.1016/j.bcdf.2021.100263. [DOI] [Google Scholar]
  41. Seera SDK, Kundu D, Gami P, Naik PK, Banerjee T. Synthesis and characterization of xylan-gelatin cross-linked reusable hydrogel for the adsorption of methylene blue. Carbohydr Polym. 2021;256:117520. doi: 10.1016/j.carbpol.2020.117520. [DOI] [PubMed] [Google Scholar]
  42. Sharma K, Morla S, Khaire KC, Thakur A, Moholkar VS, Kumar S, Goyal A. Extraction, characterization of xylan from Azadirachta indica (neem) sawdust and production of antiproliferative xylooligosaccharides. Int J Biol Macromol. 2020;163:1897–1907. doi: 10.1016/j.ijbiomac.2020.09.086. [DOI] [PubMed] [Google Scholar]
  43. Sharma K, Khaire KC, Thakur A, Moholkar VS, Goyal A. Acacia xylan as a substitute for commercially available xylan and its application in the production of xylooligosaccharides. ACS Omega. 2020;5(23):13729–13738. doi: 10.1021/acsomega.0c00896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Shi L, Dong Q, Ding K. Structure elucidation and immunomodulatory activity in vitro of a xylan from roots of Cudrania tricuspidata. Food Chem. 2014;152:291–296. doi: 10.1016/j.foodchem.2013.11.091. [DOI] [PubMed] [Google Scholar]
  45. Sluiter JB, Ruiz RO, Scarlata CJ, Sluiter AD, Templeton DW. Compositional analysis of lignocellulosic feedstocks. 1. Review and description of methods. J Agric Food Chem. 2010;58(16):9043–9053. doi: 10.1021/jf1008023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tang Q, Huang G. Improving method, properties and application of polysaccharide as emulsifier. Food Chem. 2021;376:131937. doi: 10.1016/j.foodchem.2021.131937. [DOI] [PubMed] [Google Scholar]
  47. Thompson MS, Yan TH, Saari N, Sarbini SR. A review: resistant starch, a promising prebiotic for obesity and weight management. Food Biosci. 2022 doi: 10.1016/j.fbio.2022.101965. [DOI] [Google Scholar]
  48. Ünlü CH, Kutlu M, Atıcı OG. Mannich reaction of polysaccharides: Xylan functionalization in aqueous basic medium. Carbohydr Polym. 2015;127:19–27. doi: 10.1016/j.carbpol.2015.03.024. [DOI] [PubMed] [Google Scholar]
  49. Wang S, Gao W, Wang Y, Song T, Qi H, Xiang Z. Emulsifying properties of naturally acetylated xylans and their application in lutein delivery emulsion. Carbohydr Polym. 2022;296:119927. doi: 10.1016/j.carbpol.2022.119927. [DOI] [PubMed] [Google Scholar]
  50. Wu Y, Zhang G. Synbiotic encapsulation of probiotic Latobacillus plantarum by alginate-arabinoxylan composite microspheres. LWT. 2018;93:135–141. doi: 10.1016/j.lwt.2018.03.034. [DOI] [Google Scholar]
  51. Yan J, Aznar A, Chalvin C, Birdseye DS, Baidoo EE, Eudes A, Shih PM, Loqué D, Zhang A, Scheller HV. Increased drought tolerance in plants engineered for low lignin and low xylan content. Biotechnol Biofuels. 2018;11(1):1–11. doi: 10.1186/s13068-018-1196-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zhang L, Zeng X, Qiu J, Du J, Cao X, Tang X, Sun Y, Li S, Lei T, Liu S, Lin L. Spray-dried xylooligosaccharides carried by gum Arabic. Ind Crops Prod. 2019;135:330–343. doi: 10.1016/j.indcrop.2019.04.045. [DOI] [Google Scholar]
  53. Zhang M, Zhan A, Ye Y, Liu C, Hang F, Li K, Li J. Molecular modification, structural characterization, and biological activity of xylans. Carbohydr. Polym. 2021;269:118248. doi: 10.1016/j.carbpol.2021.118248. [DOI] [PubMed] [Google Scholar]
  54. Zhang M, Bobokalonov J, Dzhonmurodov A, Xiang Z. Optimizing yield and chemical compositions of dimethylsulfoxide-extracted birchwood xylan. J Bioresour Bioprod. 2022;7(3):211–219. doi: 10.1016/j.jobab.2022.07.001. [DOI] [Google Scholar]

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