A novel glycoside hydrolase 43-like enzyme from Clostridium boliviensis is an endo-xylanase and a candidate for xylooligosaccharide production from different xylan substrates.

An uncharacterized gene encoding a glycoside hydrolase family 43-like enzyme from Clostridium boliviensis strain E-1 was identified from genomic sequence data, and the encoded enzyme, CbE1Xyn43-l, was produced in Escherichia coli. CbE1Xyn43-l (52.9 kDa) is a two-domain endo-ß-xylanase consisting of a C-terminal CBM6 and a GH43-like catalytic domain. The positions of the catalytic dyad conserved in GH43, the catalytic base (Asp74), and proton donor (Glu240) were identified in alignments including GH43-enzymes of known 3D-structure from different subfamilies. CbE1Xyn43-l is active at pH 7.0-9.0, with optimum temperature at 65°C, and a more than 7 days' half-life in irreversible deactivation studies at this temperature. The enzyme hydrolyzed birchwood xylan, quinoa stalks glucuronoarabinoxylan, and wheat arabinoxylan with xylotriose and xylotetraose as major hydrolysis products. CbE1Xyn43-l also released xylobiose from pNPX2 with low turnover (kcat of 0.044 s-1) but was inactive on pNPX, showing that a degree of polymerization of three (DP3) was the smallest hydrolyzable substrate. Divalent ions affected the specific activity on xylan substrates, which dependent on the ion could be increased or decreased. In conclusion, CbE1Xyn43-l from C. boliviensis strain E-1 is the first characterized member of a large group of homologous hypothetical proteins annotated as GH43-like and is a thermostable endo-xylanase, producing xylooligosaccharides of high DP (xylotriose and xylotetraose) producer. IMPORTANCE: The genome of Clostridium boliviensis strain E-1 encodes a number of hypothetical enzymes, annotated as glycoside hydrolase-like but not classified in the Carbohydrate Active Enzyme Database (CAZy). A novel thermostable GH43-like enzyme is here characterized as an endo-ß-xylanase of interest in the production of prebiotic xylooligosaccharides (XOs) from different xylan sources. CbE1Xyn43-l is a two-domain enzyme composed of a catalytic GH43-l domain and a CBM6 domain, producing xylotriose as main XO product. The enzyme has homologs in many related Clostridium strains which may indicate a similar function and be a previously unknown type of endo-xylanase in this evolutionary lineage of microorganisms.

Heteroxylan hydrolysis by a recombinant cellulase-free GH10 xylanase from the alkaliphilic bacterium Halalkalibacterium halodurans C-125.

The search for affordable enzymes with exceptional characteristics is fundamental to overcoming industrial and environmental constraints. In this study, a recombinant GH10 xylanase (Xyn10-HB) from the extremely alkaliphilic bacterium Halalkalibacterium halodurans C-125 cultivated at pH 10 was cloned and expressed in E. coli BL21(DE3). Removal of the signal peptide improved the expression, and an overall activity of 8 U/mL was obtained in the cell-free supernatant. The molecular weight of purified Xyn10-HB was estimated to be 42.6 kDa by SDS-PAGE. The enzyme was active across a wide pH range (5-10) with optimal activity recorded at pH 8.5 and 60 °C. It also presented good stability with a half-life of 3 h under these conditions. Substrate specificity studies showed that Xyn10-HB is a cellulase-free enzyme that conventionally hydrolyse birchwood and oat spelts xylans (Apparent Km of 0.46 mg/mL and 0.54 mg/mL, respectively). HPLC analysis showed that both xylans hydrolysis produced xylooligosaccharides (XOS) with a degree of polymerization (DP) ranging from 2 to 9. The conversion yield was 77% after 24 h with xylobiose and xylotriose as the main end-reaction products. When assayed on alkali-extracted wheat straw heteroxylan, the Xyn10-HB produced active XOS with antioxidant activity determined by the DPPH radical scavenging method (IC50 of 0.54 mg/mL after 4 h). Owing to its various characteristics, Xyn10-HB xylanase is a promising candidate for multiple biotechnological applications.

Are phenolic compounds produced during the enzymatic production of prebiotic xylooligosaccharides (XOS) beneficial: a review.

Phenolics produced during xylooligosaccharide production might inhibit xylanases and enhance the antioxidant and antimicrobial activities of XOS. The effects of phenolic compounds on xylanases may depend on the type and concentration of the compound, the plant biomass used, and the enzyme used. Understanding the effects of phenolic compounds on xylanases and their impact on XOS is critical for developing viable bioconversion of lignocellulosic biomass to XOS. Understanding the complex relationship between phenolic compounds and xylanases can lead to the development of strategies that improve the efficiency and cost-effectiveness of XOS manufacturing processes and optimise enzyme performance.

Valorization of lignocellulosic wastes for sustainable xylanase production from locally isolated Bacillus subtilis exploited for xylooligosaccharides' production with potential antimicrobial activity.

The worldwide availability of lignocellulosic wastes represents a serious environmental challenge with potential opportunities. Xylanases are crucial in lignocellulosic bio-hydrolysis, but the low enzyme productivity and stability are still challenges. In the current study, Bacillus subtilis (coded ARSE2) revealed potent xylanase activity among other local isolates. The enzyme production optimization revealed that maximum enzyme production (490.58 U/mL) was achieved with 1% xylan, 1.4% peptone, and 5% NaCl at 30 °C and pH 9. Furthermore, several lignocellulosic wastes were exploited for sustainable xylanase production, where sugarcane bagasse (16%) under solid-state fermentation and woody sawdust (2%) under submerged fermentation supported the maximum enzyme titer of about 472.03 and 485.7 U/mL, respectively. The partially purified enzyme revealed two protein bands at 42 and 30 kDa. The partially purified enzyme revealed remarkable enzyme activity and stability at 50-60 °C and pH 8-9. The enzyme also revealed significant stability toward tween-80, urea, DTT, and EDTA with Vmax and Km values of 1481.5 U/mL and 0.187 mM, respectively. Additionally, the purified xylanase was applied for xylooligosaccharides production, which revealed significant antimicrobial activity toward Staphylococcus aureus with lower activity against Escherichia coli. Hence, the locally isolated Bacillus subtilis ARSE2 could fulfill the xylanase production requirements in terms of economic production at a high titer with promising enzyme characteristics. Additionally, the resultant xylooligosaccharides revealed a promising antimicrobial potential, which paves the way for other medical applications.

Application of oligosaccharides in meat processing and preservation.

In recent decades, consumer preference and attention to foodstuff presented as healthy and with desirable nutritional information, has increased significantly. In this field, the meat industry has a challenging task since meat and meat products have been related to various chronic diseases. Functional ingredients have emerged in response to the increasing demand for healthier and more nutritious foods. On this matter, oligosaccharides such as fructooligosaccharides (FOS), xylooligosaccharides (XOS), galactooligosaccharides (GOS), and chitooligosaccharides (COS) have been presented as suitable ingredients for the meat industry with the aim of obtaining healthier meat derivatives (e.g. with low fat or sugar content, reduced amount of additives, and desirable functional properties, etc.). However, studies considering application of such oligomers in the meat sector are scarce. In addition, a large number of issues remain to be solved related both to obtaining and characterizing the oligosaccharides available in the industry and to the effect that these ingredients have on the features of meat products (mainly physicochemical and sensory). The study of new oligosaccharides, the methodologies for obtaining them, and their application to new meat products should be promoted, as well as improving knowledge about their effects on the properties of functional meat foods.

Evaluating the In Vitro and In Vivo Prebiotic Effects of Different Xylo-Oligosaccharides Obtained from Bamboo Shoots by Hydrothermal Pretreatment Combined with Endo-Xylanase Hydrolysis.

Xylo-oligosaccharides (XOS) enriched with high fractions of X2-X3 are regarded as an effective prebiotic for regulating the intestinal microflora. In this study, the original XOS solution was obtained from bamboo shoots through hydrothermal pretreatment under optimized conditions. Subsequently, enzymatic hydrolysis with endo-xylanase was performed on the original XOS solution to enhance the abundance of the X2-X3 fractions. The results demonstrated that hydrothermal pretreatment yielded 21.24% of XOS in the hydrolysate solution, and subsequent enzymatic hydrolysis significantly increased the proportion of the X2-X3 fractions from 38.87% to 68.21%. Moreover, the XOS solutions with higher amounts of X2-X3 fractions exhibited superior performance in promoting the growth of probiotics such as Bifidobacterium adolescentis and Lactobacillus acidophilus in vitro, leading to increased production of short-chain fatty acids. In the in vivo colitis mouse model, XOS solutions with higher contents of X2-X3 fractions demonstrated enhanced efficacy against intestinal inflammation. Compared with the colitis mice (model group), the XOS solution with higher X2-X3 fractions (S1 group) could significantly increase the number of Streptomyces in the intestinal microflora, while the original XOS solution (S2 group) could significantly increase the number of Bacteroides in the intestinal microflora of colitis mice. In addition, the abundances of Alcaligenes and Pasteurella in the intestinal microflora of the S1 and S2 groups were much lower than in the model group. This effect was attributed to the ability of these XOS solutions to enhance species diversity, reversing the imbalance and disorder within the intestinal microflora. Overall, this work highlights the outstanding potential of XOS enriched with high contents of X2-X3 fractions as a regulator of the intestinal microbiota and as an anti-colitis agent.

Optimization of Xylooligosaccharides Production by Native and Recombinant Xylanase Hydrolysis of Chicken Feed Substrates.

Poultry production faces several challenges, with feed efficiency being the main factor that can be influenced through the use of different nutritional strategies. Xylooligosaccharides (XOS) are functional feed additives that are attracting growing commercial interest due to their excellent ability to modulate the composition of the gut microbiota. The aim of the study was to apply crude and purified fungal xylanases, from Trichoderma harzianum, as well as a recombinant glycoside hydrolase family 10 xylanase, derived from Geobacillus stearothermophilus T6, as additives to locally produced chicken feeds. A Box-Behnken Design (BBD) was used to optimize the reducing sugar yield. Response surface methodology (RSM) revealed that reducing sugars were higher (8.05 mg/mL, 2.81 mg/mL and 2.98 mg/mL) for the starter feed treated with each of the three enzymes compared to the treatment with grower feed (3.11 mg/mL, 2.41 mg/mL and 2.62 mg/mL). The hydrolysis products were analysed by thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC) analysis and showed that the enzymes hydrolysed the chicken feeds, producing a range of monosaccharides (arabinose, mannose, glucose, and galactose) and XOS, with xylobiose being the predominant XOS. These results show promising data for future applications as additives to poultry feeds.

How Do Prebiotics Affect Human Intestinal Bacteria?-Assessment of Bacterial Growth with Inulin and XOS In Vitro.

Prebiotics are believed to exhibit high specificity in stimulating the growth or activity of a limited number of commensal microorganisms, thereby conferring health benefits to the host. However, the mechanism of action of prebiotics depends on multiple factors, including the composition of an individual's gut microbiota, and is therefore difficult to predict. It is known that different bacteria can utilize inulin and xylooligosaccharides (XOS), but an overview of which bacteria in the human gut may be affected is lacking. Detailed knowledge of how bacterial growth is affected by prebiotics is furthermore useful for the development of new synbiotics, which combine a living microorganism with a selective substrate to confer a health benefit to the host. Hence, we developed a statistical model to compare growth in vitro among typical human gut bacteria from different phylogenetic lineages. Based on continuous observation of the optical density (OD600), we compare maximal growth rates (rmax), maximal attained OD600 (ODmax), and area under the growth curve (AUC) of bacteria grown on inulin or XOS. The consideration of these three parameters suggests strain-specific preferences for inulin or XOS and reveals previously unknown preferences such as Streptococcus salivarius growth on XOS.

Autohydrolysis Application on Vine Shoots and Grape Stalks to Obtain Extracts Enriched in Xylo-Oligosaccharides and Phenolic Compounds.

Agronomic practices and the winemaking process lead to the production of considerable quantities of waste and by-products. These are often considered waste with negative effects on environmental sustainability. However, vine shoots and grape stalks can be reused, representing a potential source of xylo-oligosaccharides and polyphenols. In this context, the purpose of this work was to obtain enriched extracts using three different autohydrolysis treatments with (i) H2O, (ii) H2O:EtOH, and (iii) H2O:Amberlyst. The obtained extracts were characterized by their xylo-oligosaccharide and polyphenol profiles using LC-MS techniques. The use of ethanol during autohydrolysis allowed for greater extraction of xylan-class compounds, especially in vine shoot samples, while an increase in antioxidant activity (128.04 and 425.66 µmol TE/g for ABTS and DPPH, respectively) and in total phenol content (90.92 mg GAE/g) was obtained for grape stalks.

A sustainable process for co-production of xylooligosaccharides and ethanol from alkali treated sugarcane bagasse: A strategy towards waste management.

Present study aims at sustainable utilization of sugarcane bagasse (SCB) for production of valuable prebiotic xylooligosaccharides (XOS) along with second generation ethanol. Fractionation of SCB into hemicellulose rich liquid fraction and cellulose rich solid residue was achieved using alkaline treatment. Carbohydrate rich precipitate obtained from liquid fraction was utilized for XOS production using inhouse produced endoxylanase. XOS production from SCB xylan was optimized by employing response surface methodology. Under optimized conditions, maximum XOS yield was 227.72 mg/g of carbohydrate rich precipitates. The solid residue obtained after alkaline pretreatment was used for ethanol fermentation by prehydrolysis and simultaneous saccharification and fermentation (P-SSF) process using cellulolytic enzyme cocktail and Saccharomyces cerevisiae SM1. Maximum ethanol concentration, productivity and yield were 79.76 ± 0.16 g/L, 0.83 g/L/h and 69.38%, respectively by employing P-SSF process. Based on the experimental data it can be predicted that bioconversion of 100 g raw SCB can yield 6.26 g of XOS (DP 2-DP 5), 15.95 g ethanol and 1.44 g of xylitol. Present investigation reports an integrated process for effective bioconversion of SCB into value added products by maximum utilization of cellulosic and hemicellulosic fractions simultaneously using indigenously produced fungal enzymes.

Xylanase Production by Solid-State Fermentation for the Extraction of Xylooligosaccharides from Soybean Hulls§.

Research background: The development of a novel process for the production of xylooligosaccharides (XOS) based on the 4R concept is made possible by the integration of numerous techniques, especially enzymatic modification together with the physical pretreatment of renewable materials. This study aims to integrate the use of agricultural wastes for the production of xylanase by a new strain of Penicillium sp. and value-added products, XOS. Experimental approach: For the production of xylanase, a solid-state fermentation was performed using wheat bran as substrate. To obtain the most active crude extract of xylanase, the time frame of cultivation was first adjusted. Then, the downstream process for xylanase purification was developed by combining different membrane separation units with size exclusion chromatography. Further characterisation included determination of the optimal pH and temperature, determination of the molecular mass of the purified xylanase and analysis of kinetic parameters. Subsequently, the hydrolytic ability of the partially purified xylanase in the hydrolysis of alkali-extracted hemicellulose from soybean hulls was investigated. Results and conclusions: Our results show that Penicillium rubens produced extracellular xylanase at a yield of 21 U/g during solid-state fermentation. Using two ultrafiltration membranes of 10 and 3 kDa in combination with size exclusion chromatography, a yield of 49 % and 13-fold purification of xylanase was achieved. The purified xylanase (35 kDa) cleaved linear bonds ß-(1→4) in beechwood xylan at a maximum rate of 0.64 µmol/(min·mg) and a Michaelis constant of 44 mg/mL. At pH=6 and 45 °C, the purified xylanase showed its maximum activity. The xylanase produced showed a high ability to hydrolyse the hemicellulose fraction isolated from soybean hulls, as confirmed by thin-layer chromatography. In the hydrothermally pretreated hemicellulose hydrolysate, the content of XOS with different degrees of polymerisation was detected, while in the non-pretreated hemicellulose hydrolysate, the content of xylotriose and glucose was confirmed. Novelty and scientific contribution: Future research focusing on the creation of new enzymatic pathways for use in processes to convert renewable materials into value-added products can draw on our findings.

Production, Characterization and Prebiotic Potential of Xylooligosaccharides Produced from Wheat Bran using Enterobacter hormaechei KS1 Xylanase.

In the present investigation, xylooligosaccharides were produced from wheat bran and wheat bran extracted xylan through enzymatic hydrolysis using xylanase from novel Enterobacter hormaechei KS1. Xylooligosaccharides/reducing sugars production from wheat bran was found maximum (374 mg/g) when 4.0% of wheat bran was treated with 375 units (IU/mL) of Enterobacter hormaechei KS1 xylanase at pH 6.0 and incubated at 50 °C for 24 h of incubation. In case of wheat bran extracted xylan 419 mg/g of xylooligosaccharides were produced when 3% of extracted xylan was incubate for 8 h. Analysis of the enzymatic hydrolysate through high performance liquid chromatography equipped with refractive index detector showed the presence of xylose, xylopentose and xylohexose. The decrease in pH with 1.0% dose of xylooligosacchaides produced from extracted xylan hydrolysis using E. hormaechei KS1 xylanase showed more decrease with L. rhamnosus (6.72 to 5.94) followed by L. brevis (6.71 to 6.15) and L. plantarum (6.71 to 6.41). In case of increase in optical density both wheat bran and wheat bran extracted xylan generated xylooligosaccharides exhibited similar pattern i.e., L. rhamnosus > L. plantarum > L. brevis.

Sequence- and structure-guided improvement of the catalytic performance of a GH11 family xylanase from Bacillus subtilis.

Xylanases produce xylooligosaccharides from xylan and have thus attracted increasing attention for their usefulness in industrial applications. Previously, we demonstrated that the GH11 xylanase XynLC9 from Bacillus subtilis formed xylobiose and xylotriose as the major products with negligible production of xylose when digesting corncob-extracted xylan. Here, we aimed to improve the catalytic performance of XynLC9 via protein engineering. Based on the sequence and structural comparisons of XynLC9 with the xylanases Xyn2 from Trichoderma reesei and Xyn11A from Thermobifida fusca, we identified the N-terminal residues 5-YWQN-8 in XynLC9 as engineering hotspots and subjected this sequence to site saturation and iterative mutagenesis. The mutants W6F/Q7H and N8Y possessed a 2.6- and 1.8-fold higher catalytic activity than XynLC9, respectively, and both mutants were also more thermostable. Kinetic measurements suggested that W6F/Q7H and N8Y had lower substrate affinity, but a higher turnover rate (kcat), which resulted in increased catalytic efficiency than WT XynLC9. Furthermore, the W6F/Q7H mutant displayed a 160% increase in the yield of xylooligosaccharides from corncob-extracted xylan. Molecular dynamics simulations revealed that the W6F/Q7H and N8Y mutations led to an enlarged volume and surface area of the active site cleft, which provided more space for substrate entry and product release and thus accelerated the catalytic activity of the enzyme. The molecular evolution approach adopted in this study provides the design of a library of sequences that captures functional diversity in a limited number of protein variants.

Immobilization and Application of the Recombinant Xylanase GH10 of Malbranchea pulchella in the Production of Xylooligosaccharides from Hydrothermal Liquor of the Eucalyptus (Eucalyptus grandis) Wood Chips.

Xylooligosaccharides (XOS) are widely used in the food industry as prebiotic components. XOS with high purity are required for practical prebiotic function and other biological benefits, such as antioxidant and inflammatory properties. In this work, we immobilized the recombinant endo-1,4-ß-xylanase of Malbranchea pulchella (MpXyn10) in various chemical supports and evaluated its potential to produce xylooligosaccharides (XOS) from hydrothermal liquor of eucalyptus wood chips. Values >90% of immobilization yields were achieved from amino-activated supports for 120 min. The highest recovery values were found on Purolite (142%) and MANAE-MpXyn10 (137%) derivatives, which maintained more than 90% residual activity for 24 h at 70 °C, while the free-MpXyn10 maintained only 11%. In addition, active MpXyn10 derivatives were stable in the range of pH 4.0−6.0 and the presence of the furfural and HMF compounds. MpXyn10 derivatives were tested to produce XOS from xylan of various sources. Maximum values were observed for birchwood xylan at 8.6 mg mL−1 and wheat arabinoxylan at 8.9 mg mL−1, using Purolite-MpXyn10. Its derivative was also successfully applied in the hydrolysis of soluble xylan present in hydrothermal liquor, with 0.9 mg mL−1 of XOS after 3 h at 50 °C. This derivative maintained more than 80% XOS yield after six cycles of the assay. The results obtained provide a basis for the application of immobilized MpXyn10 to produce XOS with high purity and other high-value-added products in the lignocellulosic biorefinery field.

Natural Xylooligosaccharides Exert Antitumor Activity via Modulation of Cellular Antioxidant State and TLR4.

It has been recently proven that xylooligosaccharides (XOS) with prebiotic properties have diverse beneficial biological effects including immunomodulatory and antitumor activities. The present article focused on the chemical and biological evaluation of corn-derived commercially available XOS and aimed to elucidate their cytotoxicity and inhibitory potential against tumor cells. Spectrophotometric chemical analyses, Fourier transform infrared spectroscopy, and high-performance liquid chromatography analyses were performed. Antioxidant activity was determined by measuring the oxygen radical absorbance capacity and hydroxyl radical averting capacity. In vitro cytotoxicity assays with human cell lines derived from normal and tumor tissues, assessments of ATP production, mitochondrial membrane potential specific staining, cytokine assays, and molecular docking were used to evaluate the biological activity of XOS. The sample showed significant antioxidant activity, and it was determined that most xylose oligomers in it are composed of six units. XOS exhibited antitumor activity with pronounced inhibitory effect on lysosomes, but mitochondrial functionality was also affected. The production of proinflammatory cytokines by lipopolysaccharide-stimulated U-937 cells was reduced by XOS treatment, which suggested the involvement of Toll-like receptor 4 (TLR4)-mediated signaling in the mechanism of XOS action. Molecular docking analyses confirmed the potential inhibitory interaction between the sample and TLR4. In addition, XOS treatment had significant tumor-cell-specific influence on the glutathione antioxidant system, affecting its balance and thus contributing to the inhibition of cellular viability. The present study elucidated the tumor-inhibitory potential of commercially available XOS that could be utilized in pharmaceutical and food industry providing disease-preventive and therapeutic benefits.

Identification and Characterization of a Novel Endo-ß-1,4-Xylanase from Streptomyces sp. T7 and Its Application in Xylo-Oligosaccharide Production.

A xylanase-producing strain, identified as Streptomyces sp. T7, was isolated from soil by our lab. The endo-ß-1,4-xylanase (xynST7) gene was found in the genome sequence of strain T7, which was cloned and expressed in Escherichia coli. XynST7 belonged to the glycoside hydrolase family 10, with a molecular mass of approximately 47 kDa. The optimum pH and temperature of XynST7 were pH 6.0 and 60 °C, respectively, and it showed wide pH and temperature adaptability and stability, retaining more than half of its enzyme activity between pH 5.0 and 11.0 below 80 °C. XynST7 showed only endo-ß-1,4-xylanase activity without cellulase- or ß-xylosidase activity, and it showed maximal hydrolysis for corncob xylan in all the test substrates. Then, XynST7 was used for the production of xylo-oligosaccharides (XOSs) by hydrolyzing xylan extracted from raw corncobs. The maximum yield of the XOS was 8.61 ± 0.13 mg/mL using 15 U/mL of XynST7 and 1.5% corncob xylan after 10 h of incubation at 60 °C. The resulting hydrolysate products mainly consisted of xylobiose and xylotriose. These data indicated that XynST7 might by a promising tool for various industrial applications.

Statistical investigation of the bioprocess conditions of alkali combined twin-screw extrusion pretreatment to enhance fractionation and enzymatic hydrolysis of bulgur bran.

BACKGROUND: Bulgur bran (BB) is a potential source for the production of value-added products such as fermentable sugars and xylooligosaccharides (XOs). In this study, alkali combined twin-screw extrusion pretreatment was performed and statistically optimized to enhance fractionation and enzymatic hydrolysis of BB. The pretreatment conditions (barrel temperature, screw speed and alkali impregnation) were optimized by Box-Behnken design (BBD) to obtain the highest hemicellulose separation from BB. The obtained fractions were analyzed for the production of fermentable sugars and XOs. RESULTS: The results revealed that twin-screw extrusion of BB performed at 67 °C barrel temperature and 250 rpm screw speed after alkali impregnation at 0.02 g alkali g-1 biomass concentration provided 40.4% higher hemicellulose separation yield compared to the untreated BB. Alkali combined twin-screw extrusion pretreatment increased the enzymatic hydrolysis yield of BB fourfold, whereas a 13-fold increase was achieved after the separation of hemicellulose from pretreated BB. Xylose (X1)-free xylobiose (X2) was the main product after xylanase hydrolysis of hemicellulose fraction. SEM images confirmed the morphological modifications in BB, which were in agreement with the enhanced fractionation performance and the higher enzymatic hydrolysis yield. CONCLUSION: The results of this study suggested that pretreatment by alkali combined twin-screw extrusion followed by alkali extraction could be a reliable and effective process for fractionation of BB and production of fermentable sugars and XOs. © 2022 Society of Chemical Industry.

A Comparative Analysis for the Production of Xylooligosaccharides via Enzymatic Hydrolysis from Sugarcane Bagasse and Coconut Coir.

Xylooligosaccharides are known prebiotics that various foods and feed industries can utilize. In the present study, the xylan was extracted from sugarcane bagasse and coconut coir by NaOH treatment and it was further structurally characterized by FTIR. Furthermore, Thermomyces lanuginosus VAPS-24 was used to produce endo-xylanase, which induced the production of XOS from both of these substrates. The maximum production of XOS in 8 h using 20U enzyme concentration and 2% substrate concentration was found as 10.10 mg/ml, 5.43 mg/ml for sugarcane bagasse and coconut coir, respectively. Further, the produced XOS was structurally characterized by the NMR, HPLC and FTIR analysis. This study gives a clue that the sugarcane bagasse can be used as the potent producer for the xylooligosaccharides as compared to the coconut coir and can be utilized at the industrial level. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-022-01010-3.

A Rapid and Mild Sulfation Strategy Reveals Conformational Preferences in Therapeutically Relevant Sulfated Xylooligosaccharides.

Although sulfated xylooligosaccharides are promising therapeutic leads for a multitude of afflictions, the structural complexity and heterogeneity of commercially deployed forms (e. g. Pentosan polysulfate 1) complicates their path to further clinical development. We describe herein the synthesis of the largest homogeneous persulfated xylooligomers prepared to date, comprising up to eight xylose residues, as standards for biological studies. Near quantitative sulfation was accomplished using a remarkably mild and operationally simple protocol which avoids the need for high temperatures and a large excess of the sulfating reagent. Moreover, the sulfated xylooligomer standards so obtained enabled definitive identification of a pyridinium contaminant in a sample of a commercially prepared Pentosan drug and provided significant insights into the conformational preferences of the constituent persulfated monosaccharide residues. As the spatial distribution of sulfates is a key determinant of the binding of sulfated oligosaccharides to endogenous targets, these findings have broad implications for the advancement of Pentosan-based treatments.

Xylooligosaccharides induce stomatal closure via salicylic acid signaling-regulated reactive oxygen species and nitric oxide production in Arabidopsis.

Xylooligosaccharides (XOS) are the major coproducts of biofuel production and the most representative functional sugar enhancing animal physiology. However, little is known regarding the biological relevance of XOS to plants. Here, we found XOS triggered stomatal closure in Arabidopsis in a dose-dependent manner. Pamarcological data showed that XOS-induced stomatal closure was markedly inhibited by catalase (CAT, a reactive oxygen species [ROS] scavenger), salicylhydroxamic acid (SHAM, a peroxidase inhibitor), and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO, a nitric oxide [NO] scavenger). Moreover, XOS induced the production of ROS and NO in guard cells of Arabidopsis. ROS production was strongly restricted by CAT and SHAM, but was unaffected by treatment with diphenyleneiodonium chloride (DPI, an NADPH oxidase inhibitor) or cPTIO. NO production was suppressed by CAT, SHAM, and cPTIO, but not by DPI. The elevation of ROS level mediated by SHAM-sensitive peroxidases occurred upstream of NO. Additionally, XOS-triggered stomatal closure and ROS and NO accumulation were significantly impaired in npr1 (salicylic acid signaling) mutant plants, but were not in jar1 (jasmonic acid signaling) or ein2 (ethylene signaling) mutant plants. Furthermore, XOS-induced stomatal closure was unaffected in both ost1 and atrbohD atrbohF (abscisic acid [ABA] signaling) mutant plants. Therefore, these results indicated that the biotic sugar, XOS, can elicit stomatal closure via salicylic acid signaling-mediated production of ROS and NO, in a manner independent of ABA signaling.