Production Of Xylooligosaccharides Research Articles

Sugarcane bagasse derived xylooligosaccharides produced by an arabinofuranosidase/xylobiohydrolase from Bifidobacterium longum in synergism with xylanases

Arabinoxylan is a major hemicellulose in the sugarcane plant cell wall with arabinose decorations that impose steric restrictions on the activity of xylanases against this substrate. Enzymatic removal of the decorations by arabinofuranosidases can allow a more efficient arabinoxylan degradation by xylanases. Here we produced and characterized a recombinant Bifidobacterium longum arabinofuranosidase from glycoside hydrolase family 43 (BlAbf43) and applied it, together with GH10 and GH11 xylanases, to produce xylooligosaccharides (XOS) from wheat arabinoxylan and alkali pretreated sugarcane bagasse. The enzyme synergistically enhanced XOS production by GH10 and GH11 xylanases, being particularly efficient in combination with the latter family of enzymes, with a degree of synergism of 1.7. We also demonstrated that the enzyme is capable of not only removing arabinose decorations from the arabinoxylan and from the non-reducing end of the oligomeric substrates, but also hydrolyzing the xylan backbone yielding mostly xylobiose and xylose in particular cases. Structural studies of BlAbf43 shed light on the molecular basis of the substrate recognition and allowed hypothesizing on the structural reasons of its multifunctionality.

A fast, reliable, low‐cost, and efficient xylan extraction for xylooligosaccharides production

AbstractA fast, reliable, low‐cost, and efficient method for the recovery of xylan from agro‐residue materials is needed to improve the use of xylan in industrial processes. Xylan was extracted from two agrowastes (corncobs and wheat bran) using a cost‐effective alkali extraction method. The characteristic band patterns and bending vibrations associated with hemicellulose and beta glycoside linkage could be seen in the corncob‐extracted xylan (CCEx) and wheatbran‐extracted xylan (WBEx). Scanning electron microscopy (SEM) analysis showed that the extracted xylans consisted of aggregated and nonaggregated particles with irregular morphology and rough surfaces. Particle‐size analysis displayed average sizes of 205.6, 210.8, and 794.2 nm for commercial beechwood xylan (Bx), CCEx, and WBEx, respectively. Alkali extraction was performed with different concentrations of NaOH, from 5% to 20%, with a solid–liquid ratio of 1:20. It was found that 82.90 and 76.77% of xylan were extracted from corncob and wheat bran, respectively, which represents a higher yield obtained in a shorter duration in comparison with studies in the literature. The enzymatic hydrolysis of wheat bran and corncob that used 250 IU/mL of Enterobacter hormaechie KS1 xylanase yielded 334.38 and 162.35 mg/g of xylooligosaccharides. Their properties were studied and were found to be very similar to those of commercial xylan. They could reduce costs if applied in industry.

Comparison of lactic and propionic acid hydrolysis for production of xylo-oligosaccharides and ethanol from polysaccharides in Toona sinensis branch

Xylan-type hemicellulose hydrolysis by an organic acid solution for the production of xylo-oligosaccharides (XOS) is efficient and eco-friendly, but the effects of different organic acids on XOS production from Toona sinensis branch (TB) biomass is limited. In this work, under the conditions of 170 °C for 60 min, 33.1 % and 38.7 % XOS yields were obtained from polysaccharides present in TB by 2 % lactic acid (LA) and 6 % propionic acid (PA), respectively. Then 77 % of the lignin was removed by hydrogen peroxide−acetic acid pretreatment system, and 39.5 % and 44.7 % XOS yield were obtained from polysaccharides in delignification TB by 2 % LA and 6 % PA, respectively. It was found that PA hydrolysis, especially from delignified TB, resulted in higher XOS yield and purity compared to LA hydrolysis. Moreover, the content of byproducts (xylose, hydroxymethyl-furfural and furfural) in PA hydrolysate was lower. Following the hydrolysis process, the simultaneous saccharification and fermentation of the TB solid residue achieved an ethanol yield of 71.5 %. This work proposes an integrated process to preferentially converts the TB hemicellulose into valuable XOS and then converts the cellulose into ethanol. This process had the advantage of eliminating the need for isolation and purification, and the potential to obtain multiple products from the same raw material.

Direct enzymatic hydrolysis of solid wheat straw with endo-xylanases: Effect of the temperature on the hemicellulose release and the product profile modulation

Prebiotics are non-digestible compounds that promote intestinal microbiota growth and/or activity. Xylooligosaccharides (XOS) are new prebiotics derived from the hemicellulose fraction of lignocellulosic materials. Challenges in using those materials as sources for prebiotic compounds lie in the hemicellulose extraction efficiency and the safety of those ingredients. In this sense, this work aims to optimize hemicellulose extraction and XOS production through direct enzymatic hydrolysis of alkali pre-treated wheat straw without undesired byproducts. By increasing the temperature of the enzymatic step from 40 °C to 65 °C we achieved an improvement in the extraction yield from 55 % to 80 %. Products with different degrees of polymerization were also noticed: while XOS ≤ X6 where the main products at 40 °C, a mixture of long arabinoxylan derived polymers (ADPo) and XOS ≤ X6 was obtained at 65 °C, irrespective of the extraction yield. Thus, a modulatory effect of temperature on the product profile is suggested here. Among the XOS ≤ X6 produced, X2-X3 were the main products, and X4 was the minor one. At the end of the hydrolysis, 146.7 mg XOS per gram of pre-treated wheat straw were obtained.

Synergistic microwave and acidic deep eutectic solvent-based pretreatment of Theobroma cacao pod husk biomass for xylooligosaccharides production

The bioconversion of lignocellulosic biomass into novel bioproducts is crucial for sustainable biorefineries, providing an integrated solution for circular economy objectives. The current study investigated a novel microwave-assisted acidic deep eutectic solvent (DES) pretreatment of waste cocoa pod husk (CPH) biomass to extract xylooligosaccharides (XOS). The sequential DES (choline chloride/citric acid, molar ratio 1:1) and microwave (450W) pretreatment of CPH biomass was effective in 67.3% xylan removal with a 52% XOS yield from total xylan. Among different XOS of varying degrees of polymerization, a higher xylobiose content corresponding to 69.3% of the total XOS (68.22 mg/g CPH) from liquid fraction was observed. Enzymatic hydrolysis of residual xylan from pretreated CPH biomass with low commercial xylanase (10 IU/g) concentration yielded 24.2% XOS. The MW-ChCl/citric acid synergistic pretreatment approach holds great promise for developing a cost-effective and environmentally friendly method contributing to the sustainable production of XOS from agricultural waste streams.

A sequential bioconversion-purification process to produce high purity xylo-oligosaccharides (XOS) from Nothophagus pumilio sawdust wastes

Xylo-oligosaccharide (XOS) production from lignocellulosic biomass (LCB) is an ongoing technology in which the best raw material, catalyst, and reaction conditions remain unclear. This study proposed a three-step sequential operation to produce prebiotic Xylo-oligosaccharide (XOS) from lignocellulosic biomass (LCB) Nothophagus pumillio sawdust. First, a commercial (Enzeco Xylanase S) and recombinant (GtXyl10A) xylanase were evaluated in XOS production from pre-extracted xylan. Then, the effect of auxiliary enzyme addition (α-glucuronidase) was evaluated in xylan hydrolysis, and finally, XOS were purified by three steps of membrane filtrations: an ultrafiltration (UF) followed by two nanofiltrations (NF). From the above, XOS production yield reach 333 [mgxos∙gxylan−1] of highly purified XOS (95 %) when the process was carried out without auxiliary enzyme addition and 473 [mgxos∙gxylan−1] of 96 % purified XOS when auxiliary enzyme was added, resulting in an improvement of the XOS production yield in 42 %.This shows the feasibility to produce high purity prebiotics from sawdust wastes of the forestry industry.

Xylooligosaccharides from lignocellulosic biomass and their applications as nutraceuticals: a review on their production, purification, and characterization.

Xylooligosaccharides (XOS) are considered a potent source of prebiotics for humans. The global prebiotic market is expanding in size, was valued at USD 6.05 billion in 2021, and is expected to grow at a 14.9% compound annual growth rate (CAGR) between 2022 and 2030, indicating a huge demand. These XOS are non-digestible pentose sugar oligomers comprising mainly xylose. Xylose is naturally present in the lignocellulosic biomass (LCB), fruits, and vegetables. Apart from the prebiotic effect, these XOS have been reported to reduce blood cholesterol, antioxidant effect, increase calcium absorption, reduction of colon cancer risk, and benefit diabetic patients. Primary use of XOS is reported in the feed industry followed by health, medical use, food, and drinks. LCB mainly contains glucan, xylan, and lignin. After glucan, xylan is the second-highest available sugar on the globe composed of xylose. Therefore, the xylan fraction of LCB has great significance in producing food, feed, and energy. Glucan has been exploited for the commercial production of ethanol, xylitol, furfural, hydroxymethyl furfural, and glucose. Whereas as of now xylan has limited applications. Therefore, xylan can be exploited to convert to XOS. Production of XOS from LCB fraction not only helps to produce these at a very low price but also helps in the reduction of greenhouse gases. Its use in food and drinks is increasing as it can be derived from the abundantly and cheaply available LCB. The article provides review on production, purification, and characterization of XOS in view of their use as nutraceuticals. This article is protected by copyright. All rights reserved.

Eucalyptus grandis Forestry Residue Valorization: Distinct and Integrated Pretreatment Methods for Enhanced Xylooligosaccharide Production

AbstractEucalyptus branches and bark represent highly abundant and available feedstocks with great potential for obtaining bio-based products. Distinct and integrated pretreatment fractionation strategies for eucalyptus branches and bark were performed for the efficient production of xylooligosaccharides (XOS). By combining pretreatments, a high yield of XOS was obtained from eucalyptus branches and bark. The branches and bark were presoaked in 8% (w/w) sodium hydroxide at 60 °C for 30 min to provide a deacetylation effect. The residues were then hydrothermally treated. The findings revealed that 4.64% of XOS originated from the bark and 6.19% from eucalyptus branches. It has been demonstrated that xylan may be selectively depolymerized during pretreatment by preventing excessive hydrolysis through the use of deacetylation in the first phase of the process. More XOS was produced using hydrothermal treatment, yielding 8.00% (w/w) in the branches and 5.12% in the bark. A significant amount of XOS with DP 2–5 might be obtained in certain experiments, up to 60%, but the most abundant XOS are usually those with DP > 5 (approximately 80% of all XOS). This work provides new insights into the effective generation of XOS under relatively mild conditions by overcoming the recalcitrant structure of eucalyptus branches and bark, representing a noteworthy advancement towards forestry leftover valorization.

Two-domain GH30 xylanase from human gut microbiota as a tool for enzymatic production of xylooligosaccharides: Crystallographic structure and a synergy with GH11 xylosidase

Production of value-added compounds and sustainable materials from agro-industrial residues is essential for better waste management and building of circular economy. This includes valorization of hemicellulosic fraction of plant biomass, the second most abundant biopolymer from plant cell walls, aiming to produce prebiotic oligosaccharides, widely explored in food and feed industries. In this work, we conducted biochemical and biophysical characterization of a prokaryotic two-domain R. champanellensis xylanase from glycoside hydrolase (GH) family 30 (RcXyn30A), and evaluated its applicability for XOS production from glucuronoxylan in combination with two endo-xylanases from GH10 and GH11 families and a GH11 xylobiohydrolase. RcXyn30A liberates mainly long monoglucuronylated xylooligosaccharides and is inefficient in cleaving unbranched oligosaccharides. Crystallographic structure of RcXyn30A catalytic domain was solved and refined to 1.37 Å resolution. Structural analysis of the catalytic domain releveled that its high affinity for glucuronic acid substituted xylan is due to the coordination of the substrate decoration by several hydrogen bonds and ionic interactions in the subsite −2. Furthermore, the protein has a larger β5-α5 loop as compared to other GH30 xylanases, which might be crucial for creating an additional aglycone subsite (+3) of the catalytic site. Finally, RcXyn30A activity is synergic to that of GH11 xylobiohydrolase.

Benzoic acid catalyzed production of xylose and xylooligosaccharides from poplar

The proficient synthesis of xylose and xylooligosaccharides (XOS) from lignocellulosic biomass proved to be a formidable undertaking, particularly in the absence of preliminary hemicellulose isolation. In this study, the potential of benzoic acid (BZA), a non-toxic and edible acidic catalyst, was assessed for the pretreatment of poplar in order to produce xylose and xylooligosaccharides (XOS). At low temperatures (< 160°C) or low acid concentrations (< 2%), the main product obtained from the depolymerization of hemicellulose was XOS, and no significant depolymerization occurred on a large scale. The results showed that the highest extraction yield of hemicellulose was 93.1% with 2% BZA at 160°C for 0.75 h, hemicellulose was mainly converted into xylose and XOS, with xylose accounting for 27.1% and XOS accounting for 46.7%. Meanwhile, the results of the process simulation of industrial energy consumption indicated that the amount of water added was the main factor affecting energy consumption in industrialization, followed by temperature. This study presented a promising approach for the industrial-scale manufacturing of xylose and XOS from poplar as a primary resource.

Valorization of agro-wastes by Bacillus altitudinis XYL17 through simultaneous production of xylanase, xylooligosaccharides, and antioxidant compounds

Bacterial strains were isolated from undisturbed regions of Sikkim Himalaya for the production of highly active xylanase and xylooligosaccharides (XOS). Seven bacterial isolates showed xylanase production while fermenting beechwood xylan (BX) and the agrowastes corncob (CC), rice straw (RS), and sugarcane bagasse (SB). Bacillus altitudinis XYL17, isolated from soil at an elevation of 4083 m, produced the highest amount of xylanase. Maximum xylanase production was observed during BX fermentation after 24 h with the an observed activity of 6576.96 ± 82.75 U/mg. CC was observed to be the best agrowaste substrate for xylanase production with an observed activity of 4150.25 ± 94.64 U/mg. Xylopentaose was the most prominent XOS, with a highest observed content of 11.206 ± 0.331 mg/mL in RS hydrolysate. Significant increase of total phenolic content was observed for hydrolysates of B. altitudinis XYL17 fermented agro-wastes as compared to the control, while there was no significant difference (p < 0.05) in phenolic content between the control and fermented BX. The biotechnological potential of the XOS-rich hydrolysate was demonstrated by the observed radical scavenging, HOCl reducing, and myeloperoxidase (MPO) inhibitory activity. MPO inhibitory IC50 of 6.88 mg/mL and 9.4 mg/mL was observed for hydrolysates of SB and CC, respectively. We conclude that B. altitudinis XYL17 can be used for the valorization of agricultural wastes for the simultaneous production of highly efficient xylanase along with XOS and other antioxidant compounds.

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

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-β-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.

Production of Lipid and Xylo-oligosaccharides from Cellulose and Hemicellulose in Reed Sawdust

Production of Lipid and Xylo-oligosaccharides from Cellulose and Hemicellulose in Reed Sawdust

Sustainable production of cellulose and hemicellulose-derived oligosaccharides from pineapple leaves: Impact of hydrothermal pretreatment and controlled enzymatic hydrolysis

Globally, the demands for sustainably sourced functional foods like prebiotic oligosaccharides have been constantly increasing. This study assessed the potential of pineapple leaves (PL) as lignocellulosic feedstock for sustainable production of cellulose and hemicellulose-derived oligosaccharides through its hydrothermal pretreatment (HT) followed by controlled enzymatic hydrolysis. PL was subjected to HT at 160, 175, and 190 °C for 20, 30, 60, and 90 min without any catalyst for xylooligosaccharide (XOS) production, whereas, the resulting solid content after HT was subjected to controlled enzymatic hydrolysis by commercial cellulase using conduritol B epoxide (0.5–5 mM) for glucooligosaccharides (GOS) production. HT at 160 °C for 60 min resulted in maximum yield of XOS and GOS at 23.7 and 18.3 %, respectively, in the liquid phase. Controlled enzymatic hydrolysis of HT treated (160 °C) PL solids for 20 and 30 min yielded ∼ 174 mg cellobiose/g dry biomass within 24 h, indicating overall high oligosaccharide production.

Valorisation of sawdust-based spent mushroom substrate for sustainable xylooligosaccharides production using low-cost crude xylanases from Aspergillus flavus KUB2

ABSTRACT Spent mushroom substrate (SMS), a lignocellulosic waste after mushroom production is generally discarded without proper management. There is increasing interest in the sustainable transformation of lignocellulosic waste into high-value products. Within this context, the present study investigated the potential of the SMS from the cultivation of Pleurotus pulmonarius and Auricularia auricula on rubber tree wood sawdust as substrates for xylooligosaccharides (XOS) production. SMS samples from these two edible mushrooms were extracted using alkaline xylan extraction, producing maximum true recovery amounts of xylan in the range 34.61%–37.49% using 18% NaOH at 70 °C for 3 h. Production of XOS from alkaline-extracted xylan from the SMS samples of both mushroom species using economically crude xylanases from Aspergillus flavus KUB2 resulted in XOS (X2–X5) production of 241.47–249.04 mg/g, with X3 as the predominant XOS product. The produced XOS had excellent prebiotic activity and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and contained high total phenolic contents. The combined beneficial bioactivities in terms of prebiotic and antioxidant properties suggested that the XOS produced from sawdust-based SMS samples of P. pulmonarius and A. auricula could be promising ingredients for both food and pharmaceutical applications.

Novel approach for corn straw biorefineries: Production of xylooligosaccharides, lignin and ethanol by nicotinic acid hydrolysis and pentanol pretreatment

The productive separation and conversion of corn straw offers significant prospects for the economic viability of biorefineries centered on straw resources. In this work, a graded utilization method was proposed to produce xylo-oligosaccharides (XOS), ethanol and lignin from corn straw by nicotinic acid (NA) hydrolysis and water/pentanol pretreatment. A XOS yield of 52.6 % was achieved under optimized conditions of 100 mM NA, 170 °C and 30 min. The solid residue was directly treated with water/pentanol, achieving a lignin removal rate of 79.7 %, and the total XOS yield was improved to 62.6 %. The lignin recovered from pentanol had a high purity of 97.6 %, with high phenolic OH content. Simultaneous saccharification and fermentation of final residue resulted in an ethanol yield of 92.0 %, which yielded 55.3 g/L ethanol. Thus, NA hydrolysis and water/pentanol pretreatment provided an efficient, environmentally friendly approach to fractionate corn straw for the co-production of XOS, ethanol, and lignin.

An eco-friendly process for low-degree polymerization xylo-oligosaccharides production from corncob via enzyme adsorption-hydrolysis integrated approach by reconstructing fusion protein

Xylo-oligosaccharides (XOS) have gained significant attention for their extensive health-boosting characteristics. However, the tedious procedures involved in lignocellulose pretreatment and XOS purification, coupled with the low efficiency of enzymatic hydrolysis, constrain the broader commercialization of XOS. In this study, the effects of fusion order and linker composition on enzyme activity and thermostability of fusion proteins consisting of a GH11 xylanase mutant (XYNm) and an expansin (EXLX) were investigated. The results indicated that the reconstructed protein XYNm-R2-EXLX exhibited 2.6-fold activity higher than EXLX-R2-XYNm. Furthermore, the denaturation thermodynamics evaluation demonstrated a significant enhancement in the thermostability of XYNm-R2-EXLX. Additionally, the reconstructed fusion protein showed a notable affinity adsorption efficiency on corncob, reaching a remarkable 95.5%. Subsequently, the fusion enzyme-corncob complex underwent direct hydrolysis, and a yield of 58.7 g XOS/kg corncob was obtained by employing RSM optimization techniques. Finally, a membrane-assisted refining process was implemented, resulting in the recovery of 68.2% (w/w) of the initial XOS (X2-X4) with a purity of 96.2%. Here, a chemical free and eco-friendly bioprocess for production of low-degree polymerization XOS was developed, eliminating the need for xylan separation from corncob and simplifying the process for product purification. These findings exhibited the great potential in XOS (X2-X4) production.

Multi-point immobilization of GH 11 endo-β-1,4-xylanase on magnetic MOF composites for higher yield of xylo-oligosaccharides

GH 11 endo-β-1,4-xylanase (Xy) was a crucial enzyme for xylooligosaccharides (XOS) production. The lower reusability and higher cost of purification has limited the industrial application of Xy. Addressing these challenges, our study utilized various immobilization techniques, different supports and forces for Xy immobilization. This study presents a new method in the development of Fe3O4@PDA@MOF-Xy which is immobilized via multi-point interaction forces, demonstrating a significant advancement in protein loading capacity (80.67 mg/g), and exhibiting remarkable tolerance to acidic and alkaline conditions. This method significantly improved Xy reusability and efficiency for industrial applications, maintaining 60 % activity over 10 cycles. Approximately 23 % XOS production was achieved by Fe3O4@PDA@MOF-Xy. Moreover, the yield of XOS from cobcorn xylan using this system was 1.15 times higher than that of the free enzyme system. These results provide a theoretical and applicative basis for enzyme immobilization and XOS industrial production.

Efficient production of xylooligosaccharides from Camellia oleifera shells pretreated by pyruvic acid at lower temperature

XOS production from lignocellulose using organic carboxylic acids and alkyd acids has been widely reported. However, it still faces harsh challenges such as high energy consumption, high cost, and low purity. Pyruvic acid (PYA), a carbonyl acid with carbonyl and carboxyl groups, was used to produce XOS due to its stronger catalytic activity. In this work, XOS was efficiently prepared from COS in an autoclave under the condition of 0.21 M PYA-121 °C-35 min. The total yield of XOS reached 68.72 % without producing any toxic by-products, including furfural (FF) and 5-hydroxymethylfurfural (5-HMF). The yield of xylobiose (X2), xylotriose (X3), xylotetraose (X4), and xylopentaose (X5) were 20.58 %, 12.47 %, 15.74 %, and 10.05 %, respectively. Meanwhile, 89.05 % of lignin was retained in the solid residue, which provides a crucial functional group for synthesizing layered carbon materials (SRG-a). It achieves excellent electromagnetic shielding (EMS) performance through graphitization, reaching −30 dB at a thickness of 2.0 mm. The use of a PYA catalyst in the production of XOS has proven to be an efficient method due to lower temperature, lower acid consumption, and straightforward operation.

Production of xylooligosaccharides from different pretreated Camellia oleifera shell with crude enzyme from Aspergillus fumigatus

Production of xylooligosaccharides (XOS) from Camellia oleifera shell (COS) using the crude enzyme from Aspergillus fumigatus (A. fumigatus) was investigated. COS was subjected to different pretreatments, namely thermal autoclaving, combination of diluted acid, diluted alkali or concentrated alkali with thermal autoclaving, microwave-assited ethanol pretreatment. The results showed that the combination of diluted alkali with thermal autoclaving is the best pretreatment for XOS production. The optimal conditions for enzymatic xylooligosaccharides production from the best pretreatment are: temperature 50 °C, pH 4.5, substrate concentration 8% (m/V), enzymatic hydrolysis time 9 h, and enzyme dose of 40 IU/g substrate. A maximum yield of 17.08 ± 0.67 g/L of XOS was obtained under the above optimum conditions for the best pretreatment.