Production Of Xylooligosaccharides Research Articles

Carbohydrates valorization of Quinoa (Chenopodium quinoa) stalk in xylooligosaccharides and carotenoids as emergent biomolecules

Quinoa (Chenopodium quinoa) stalk (QS) is an unvalued and renewable lignocellulosic waste. Aiming to its valorization, autohydrolysis process (178 °C and 10 bar) for xylooligosaccharides (XOS) production and enzymatic hydrolysate of residual solid for carotenoids production by Rhodotorula glutinis were evaluated. The highest yield of XOS (79 mg/g of biomass) containing mainly xylotetraose (X4) to xylohexaose (X6) was achieved in 20 min of process, while high xylobiose (X2, 18 mg/g of QS) and xylotriose (X3, 18 mg/g of QS) contents were observed in 45 min. In ultrafiltration-UF using membrane with nominal MWCO of 10 kDa, higher permeate flux was observed at pH 6.6 than at lower pH values (3.6, 4.6, and 5.6). In the retentate after nanofiltration-NF (150–300 Da), 56 % of total XOS (X2-X6) was recovered. Finally, 4.1 mg of carotenoids/g of dry cells weight, corresponding to 6.27 mg of carotenoids/g of total sugar consumed, were produced by the yeast using residual solids hydrolysate. Therefore, quinoa stalk was shown as an interesting source for high value biomolecules production.

Green production of prebiotic xylooligosaccharides via enzymatic hydrolysis from xylan black liquor of oil palm frond

The black liquor generated from the alkaline pretreatment of lignocellulosic biomass is usually disposed of into wastewater, which could lead to environmental pollution. Alkaline black liquor (ALBL) contains a large amount of xylan with a small fraction of lignin, making it a promising raw material for the production of xylooligosaccharides (XOS). In this study, xylan was extracted from the ALBL generated upon treating the oil palm frond (OPF) with sodium hydroxide via two-stage precipitation for the separation of lignin and recovery of xylan. As a result, approximately 84.0% of xylan retrieved from the ALBL was recovered. Subsequently, enzymatic hydrolysis was optimized to recover the maximum amount of XOS from xylan. The results showed that enzymatic hydrolysis produced the highest XOS (62.5%) under optimal conditions of 50 °C, 4 U/mL xylanase, and 3% xylan loading for 48 h. The study provides insight for maximizing utilization of ALBL of OPF for future biorefinery economy.

Enzymatic Cocktail Formulation for Xylan Hydrolysis into Xylose and Xylooligosaccharides.

In the context of a biorefinery, lignocellulosic materials represent an important source of raw material for the bioconversion of cellulose, hemicellulose, and lignin into value-added products, such as xylose for fermentation, oligosaccharides, and bioplastics for packaging. Among the most abundant lignocellulosic materials in Brazil, sugarcane bagasse biomass stands out, as it is rich in cellulose and hemicellulose. In this context, through an experimental design, this study developed a robust enzyme cocktail containing xylanases and accessory enzymes to complete the hydrolysis of xylan from sugarcane bagasse, obtaining a low xylose yield and concentration (9% and 1.8 g/L, respectively, observed in experiment number 16 from the complete hydrolysis of a xylan assay), a fermentable sugar that is important in the production of second-generation ethanol, and a high xylooligosaccharides (XOS) yield and concentration (93.1% and 19.6 g/L, respectively, obtained from a xylooligosaccharides production assay); in general, xylan has prebiotic activities that favor an improvement in intestinal functions, with immunological and antimicrobial actions and other benefits to human health. In addition to completely hydrolyzing the sugarcane bagasse xylan, this enzymatic cocktail has great potential to be applied in other sources of lignocellulosic biomass for the conversion of xylan into xylose and XOS due to its enzymes content, involving both main chain and pendant groups hydrolysis of hemicelluloses.

Breeding of a thermostable xylanase-producing strain of Myceliophthora thermophila by atmospheric room temperature plasma (ARTP) mutagenesis.

Introduction: Hemicellulose is an important component in lignocellulose materials, which is second only to cellulose, accounting for 15%-35% of the dry weight of plants. In the current situation of energy shortage, making full use of lignocellulose materials to produce fuel ethanol has become an important way to solve the energy problem. Xylanase plays a crucial role in the utilization of hemicellulose. It is a necessary means to reduce the cost of hemicellulose utilization by improving the activity of xylanase. Moreover, most naturally xylanases are mesophilic enzymes, which limits their industrial application. Methods:In this study, Myceliophthora thermophila was used to produce xylanases and a thermostable mutant M 2103 was obtained by atmospheric room temperature plasma (ARTP) mutagenesis. The research work started with exploring the effects of ARTP mutagenesis on the antioxidase system [superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), polyphenol oxidase (PPO), and antioxidant capacity (AOC)] of M. thermophile, and found that superoxide dismutase activity increased by 221.13%, and polyphenol oxidase activity increased by 486.04% as compared with the original strain when the implantation time was 300s. So as to determine the conditions for subsequent mutagenesis. Results and Discussion:For the mutant M 2103, the reaction temperature for xylanase production remained stable in the range of 70°C-85°C. Its optimum temperature was 75°C, which was 15°C higher than that of the original strain. And its xylanase activity increased by 21.71% as compared with the original strain. M 2103 displayed a significantly higher relative xylanase activity than the original strain in the acidic (pH 4.0-7.0) range, and the xylanase activity was relatively stable in the pH range of 6.0-8.5. These results provide an alternative biocatalyst for the production of xylooligosaccharide, and a potential usage of ARTP in the mutagenesis of thermostable mutant.

Xylopentose production from crop residue employing xylanase enzyme

To produce xylo-oligosaccharides (XOS) from the agriculture waste, which included, green coconut and vegetable cocktail. The two pretreatment – hydrogen peroxide-acetic acid (HP-AC) and sodium hypochlorite-sodium hydroxide (SH-SH) – were used for this study. The optimal conditions for the pretreatment were 80 °C, 4.0 % NaClO, and 2 h, followed by 0.08 % NaOH, 55 °C, and 1 h. Further enzymatic hydrolysis of green coconut (GC) and vegetable cocktail (VC) were performed and found in case of GC, the best outcomes were observed. Different types of XOS were obtained from the treated biomass whereas a single type of XOS xylo-pentose was obtained in high quantity (96.44 % and 93.09 % from CG and VC respectively) with the production of other XOS < 2 %. This study presents a reasonably secure and economical method for turning secondary crop residue into XOS and fermentable sugars.

Application of Endoxylanases of Bacillus halodurans for Producing Xylooligosaccharides from Empty Fruit Bunch

Endo-1,4-β-xylanase catalyzes the random hydrolysis of β-1,4-D-xylosidic bonds in xylan, resulting in the formation of oligomers of xylose. This study aims to demonstrate the promise of endoxylanases from alkaliphilic Bacillus halodurans for the production of xylooligosaccharides (XOS) from oil palm empty fruit bunch (EFB) at high pH. Two enzyme preparations were employed: recombinant endoxylanase Xyn45 (GH10 xylanase) and nonrecombinant endoxylanases, a mixture of two extracellular endo-1,4-β-xylanases Xyn45 and Xyn23 (GH11 xylanase) produced by B. halodurans. EFB was first treated with an alkaline solution. Then, the dissolved xylan-containing fraction was retained, and a prepared enzyme was added to react at pH 8 to convert xylan into XOS. Compared with the use of only Xyn45, the combined use of Xyn45 and Xyn23 resulted in a higher yield of XOS, suggesting the synergistic effect of the two endoxylanases. The yield of XOS obtained from EFB was as high as 46.77% ± 1.64% (w/w), with the xylobiose-to-xylotriose ratio being 6:5. However, when the enzyme activity dose was low, the product contained more xylotriose than xylobiose. Four probiotic lactobacilli and bifidobacteria grew well on a medium containing XOS from EFB. The presence of XOS increased cell mass and reduced pH, suggesting that XOS promoted the growth of probiotics.

Topochemical Design of Cellulose-Based Carriers for Immobilization of Endoxylanase

Xylooligosaccharides (XOSs) gained much attention for their use in food and animal feed, attributed to their prebiotic function. These short-chained carbohydrates can be enzymatically produced from xylan, one of the most prevalent forms of hemicellulose. In this work, endo-1,4-β-xylanase from Thermotoga maritima was immobilized on cellulose-based beads with the goal of producing xylooligosaccharides with degrees of polymerization (DPs) in the range of 4-6 monomeric units. More specifically, the impact of different spacer arms, tethers connecting the enzyme with the particle, on the expressed enzymatic activity and oligosaccharide yield was investigated. After surface functionalization of the cellulose beads, the presence of amines was confirmed with time of flight secondary ion mass spectrometry (TOF-SIMS), and the influence of different spacer arms on xylanase activity was established. Furthermore, XOSs (DPs 2-6) with up to 58.27 mg/g xylan were obtained, which were greatly enriched in longer oligosaccharides. Approximately 80% of these XOSs displayed DPs between 4 and 6. These findings highlight the importance of topochemical engineering of carriers to influence enzyme activity, and the work puts forward an enzymatic system focusing on the production of longer xylooligosaccharides.

Green Process for Xylooligosaccharides Production using an Eucalyptus Kraft Pulp.

Xylooligosaccharides (XOS) are oligomers with recognized and important prebiotic properties, whose consumption is associated with several health benefits, including a positive impact on the immune system. In this work, XOS were produced through a green process of enzymatic hydrolysis performed directly on an intermediate product from a pulp and paper industry, Eucalyptus bleached kraft pulp. Focusing on an industrial, sustainable and more economical application, two goals were defined and validated: (i) no pretreatment of the substrate and (ii) the replacement of the commonly used buffer solution as reaction medium for only water. The influence of the most relevant operating conditions on the production of XOS as well as the respective yields obtained were very similar when using either buffer or water as the reaction medium. For the use of water, although the solution pH decreases during the enzymatic reaction, this change did not affect the production of XOS. For the optimized conditions, 80°C and 100 U/gpulp, a maximum yield of 31.4 ± 2.6% per total xylan in the pulp was obtained, resulting in more than 50kg of XOS per ton of pulp. The correspondent hydrolysate was mainly composed by xylobiose (66%) and xylotriose (29%), oligomers with the highest prebiotic effect.

Optimizing tri-acid mixture hydrolysis: An improved strategy for efficient xylooligosaccharides production from corncob

Propionic acid (PA) hydrolysis of corncob for xylooligosaccharides (XOS) production has the advantages of simple operation, high XOS yield and less by-products, but the high price of PA limits its application. Therefore, partially replacing PA with less expensive organic acids, such as formic acid (FA) and acetic acid (AC), may lower the cost of hydrolysis in XOS production. This work investigated the feasibility of XOS production from corncob using a tri-acid mixture of FA, AC and PA. A high XOS yield of 69.1 % was achieved under the optimal FA:PA:AC volume ratio of 1:5:4 at 150 °C for 50 min. Overall, in the XOS production from corncob, it was able to replace 60 % of PA with FA and AC, and decreased the hydrolysis temperature from 170 °C to 150 °C, all of which were important to lower the cost of XOS production using organic acid hydrolysis.

A New Insight into the Composition and Physical Characteristics of Corncob—Substantiating Its Potential for Tailored Biorefinery Objectives

Corncobs of four different corn varieties were physically segregated into two different anatomical portions, namely the corncob outer (CO) and corncob pith (CP). The biomass composition analysis of both the CO and CP was performed by four different methods. The CP showed a higher carbohydrate and lower lignin content (83.32% and 13.58%, respectively) compared with the CO (79.93% and 17.12%, respectively) in all of the methods. The syringyl/guaiacyl (S/G) ratio was observed to be higher in the CP (1.34) than in the CO (1.28). The comprehensive physical characterization of both samples substantiated the lower crystallinity and lower thermal stability that was observed in the CP compared to the CO. These properties make the CP more susceptible to glycanases, as evident from the enzymatic saccharification of CP carried out with a commercial cellulase and xylanase in this work. The yields obtained were 70.57% and 88.70% of the respective theoretical yields and were found to be equal to that of pure cellulose and xylan substrates. These results support the feasibility of the tailored valorization of corncob anatomical portions, such as enzymatic production of xylooligosaccharides from CP without pretreatment combined with the bioethanol production from pretreated CO to achieve an economical biorefinery output from corncob feedstock.

Xylan extraction from hardwoods by alkaline pretreatment for xylooligosaccharide production: A detailed fractionation analysis

In the last decades, the production of value-added products from lignocellulosic biomass (LCB) has gained relevance. Xylans, which are the main hemicellulose compounds in LCB, may be extracted by alkaline pretreatment and employed for xylooligosaccharide (XOS) production. However, xylan extraction currently works as a black box due to the lack of characterization of the involved streams. Therefore, the appropriate operational conditions often remain unclear, especially in hardwoods. In this study, alkaline/thermal pretreatments at different operational conditions were evaluated for xylan extractions from Chilean Nothofagus species sawdust, determining the chemical compositions of the fractions at each step of the process. Results indicated that increasing alkali concentration (NaOH) leads to a higher xylan extraction, but also to high salt production during the acid neutralization step, decreasing xylan's purity and therefore XOS production. In this context, decreasing NaOH concentration and neutralizing it by membrane filtration, allow extracting xylans (62.5 %) of higher-purity (77 %).

Production, Properties and Applications of Xylooligosaccharides (XOS): A Review

Xylooligosaccharides derived from Xylan, significant component of hemicellulose in lignocellulosic plant biomass, are essential raw materials used in the food, pharmaceutical and agricultural industries. They find application in these industries due to their unique physicochemical properties such as pH stability, growth regulatory ability, and anti-allergic property. In addition, they have been shown to exhibit promising antimicrobial, anti-cancer, antioxidant and anti-inflammatory activities. Xylooligosaccharides convert waste into valuable foods, pharmaceuticals, and agricultural products, promoting health, the economy, and the environment.

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-β-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 &gt;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.

A biorefinery approach for sequential extraction of commercial grade xylan and alkali lignin from alkali pretreated sugarcane tops hydrolysate

Sequential extraction of xylan and lignin from alkali pretreated sugarcane tops (apSCT) hydrolysate was carried out. The xylan extracted from apSCT hydrolysate (apSCTx) was 26.5 g and the alkali lignin (apSCTal) was 8.5 g from 100 g of raw SCT. The composition analysis of apSCTx by high performance liquid chromatography showed 74% D-xylose, 15% D-glucuronic acid and 11% L-arabinose residues. The analysis of apSCTx by high performance size exclusion chromatography showed molecular size, ∼58 kDa. The first derivative of thermogravimetric analysis of apSCTx showed a Td (thermal degradation temperature) at 275ºC. The elemental (CHNS) analysis of apSCTx displayed the presence of only carbon and hydrogen. The absence of nitrogen and sulfur in the extracted apSCTx displayed that the xylan is pure and without the protein and lignin. The specific activity of recombinant clostridial endoxylanase (CtXyn11A) was 1823 ± 51 U/mg. Liquid Chromatography-Mass Spectroscopy and Thin-layer chromatography analyses of CtXyn11A hydrolyzed apSCTx displayed a series of linear xylooligosaccharides ranging between 2 and 5 degrees of polymerization. apSCTx can serve as a potential commercial substrate and also for xylooligosaccharides production. The characterization of apSCTal by Field-Emission Scanning Electron Microscopy showed the exposed spherical structures of alkali lignin. Fourier Transform Infrared spectroscopic analysis of apSCTal displayed the peaks corresponding to those obtained from commercial alkali lignin. CHNS analysis of apSCTal confirmed the absence of protein impurities. The first derivative of the thermogravimetric analysis of apSCTal showed Td at 362ºC, similar to the previously reported softwood lignin. The extracted apSCTal can be used as feedstock for the production of aromatic chemicals. The biorefinery approach of sequential extraction of apSCTx and apSCTal from alkali pretreated SCT hydrolysate waste can enhance the economics of the extraction process.

Kinetic studies and predictions for xylo-oligosaccharide production by acidic hydrolysis of xylan with various acids

Acidic hydrolysis of xylan for xylooligosaccharide production has already been intensively investigated allowing for its high efficiency and simple manufacturing procedure. For the purpose of gaining insight into the relationship between the degradation process of xylan and yields of xylooligosaccharides, in this study, four acids (sulfuric, maleic, lactic and acetic) with different pKa values were employed to hydrolyze xylan and different kinetic models were established by further analyzing the hydrolysis process from the perspective of reaction rate and activation energy. All results confirmed that weaker organic acids were more suitable for the production of xylooligosaccharides, providing a higher generation rate of xylooligosaccharides and a lower xylose formation rate. A xylooligosaccharides’ yield of 47.2% could be achieved by lactic acid hydrolysis based on prediction of kinetic models. This study provides a convenient and convincing method for the selection of acid catalysts on xylooligosaccharides production from xylan.

Efficient co-production of xylooligosaccharides and glucose from lignocelluloses by acid/pentanol pretreatment: Synergetic role of lignin removal and inhibitors

A novel technology for co-production of xylooligosaccharides (XOS) and glucose from Monterey pine sawdust and wheat straw was introduced using dilute acid (DA)/pentanol pretreatment. Effects of pretreatment severity (PS), lignin removal, and inhibitors with byproduct concentrations on XOS production were investigated. Optimal identified conditions (PS: 3.71; 170 °C, 45 min) resulted in maximum XOS of 48.65 % (pine sawdust) and 46.85 % (wheat straw), due to appropriate lignin removal (pine sawdust, 88.5 %; wheat straw, 89.7 %) and formation of small amounts of inhibitors and byproducts. Enzymatic hydrolysis of optimal pretreated solid residues yielded 88.65 % and 93.34 % glucose in pine sawdust and wheat straw, respectively. Biomass characterization revealed that DA/pentanol pretreatment enhanced porosity and pore size along with removal of amorphous fractions in both samples, thereby increasing cellulose accessibility and glucose yield. This study demonstrated lignin removal and low formation of inhibitors and byproducts, effectively enhancing XOS and glucose production from lignocellulosic biomass.

Combining autohydrolysis with xylanase hydrolysis for producing xylooligosaccharides from Jiuzao

In this report, a versatile process for producing xylooligosaccharides (XOS) with great prebiotic potential from Jiuzao, a residue from Baijiu distillation, was designed. This two-stage process integrated autohydrolysis with subsequent enzymatic hydrolysis by the recombinant thermostable xylanase, XynAS, produced by Escherichia coli. The effect of several autohydrolysis parameters (i.e., temperature, time, particle size and solid-liquid ratio) on XOS yield were examined by single factor and response surface methods. The maximum XOS yield was 30.4%, which was obtained at 181.2 °C for 20 min with a 1:13.4 solid-liquid ratio for the autohydrolysis step, and pH 6, 60 °C, 5 U/mL XynAS loading and an incubation period of 2 h for the enzymatic hydrolysis step. Using these reaction conditions, xylobiose and xylotriose were the major XOS products, constituting 86.6% of the total XOS yield. The results demonstrated that autohydrolysis combined with xylanase hydrolysis is an effective and eco-friendly approach for XOS production using Jiuzao as the substrate.

Characterization of a GH5 endoxylanase from Penicillium funiculosum and its synergism with GH16 endo-1,3(4)-glucanase in saccharification of sugarcane bagasse

The production of second-generation fuels from lignocellulosic residues such as sugarcane bagasse (SCB) requires the synergistic interaction of key cellulose-degrading enzymes and accessory proteins for their complete deconstruction to useful monomeric sugars. Here, we recombinantly expressed and characterized unknown GH5 xylanase from P. funiculosum (PfXyn5) in Pichia pastoris, which was earlier found in our study to be highly implicated in SCB saccharification. The PfXyn5 has a molecular mass of ~ 55 kDa and showed broad activity against a range of substrates like xylan, xyloglucan, laminarin and p-nitrophenyl-β-d-xylopyranoside, with the highest specific activity of 0.7 U/mg against xylan at pH 4.5 and 50 °C. Analysis of the degradation products of xylan and SCB by PfXyn5 showed significant production of xylooligosaccharides (XOS) with a degree of polymerization (DP) ranging from two (DP2) to six (DP6), thus, suggesting that the PfXyn5 is an endo-acting enzyme. The enzyme synergistically improved the saccharification of SCB when combined with the crude cellulase cocktail of P. funiculosum with a degree of synergism up to 1.32. The PfXyn5 was further expressed individually and simultaneously with a notable GH16 endoglucanase (PfEgl16) in a catabolite-derepressed strain of P. funiculosum, PfMig188, and the saccharification efficiency of the secretomes from the resulting transformants were investigated on SCB. The secretome of PfMig188 overexpressing Xyn5 or Egl16 increased the saccharification of SCB by 9% or 7%, respectively, over the secretome of PfMig188, while the secretome of dual transformant increased SCB saccharification by ~ 15% at the same minimal protein concentration.

GROWTH STIMULATION OF Lactococcus lactis sps BY XYLOOLIGOSACCHARIDES PRODUCED USING SINGLE FERMENTATIVE STEP FROM LIGNOCELLULOSIC WASTE BY Massilia Timonae B2YR

As a source of xylan, lignocellulose waste such as wheat bran and aquatic weed Pistia stratiotes were employed, together with potassium nitrate as a nitrogen source, which were optimized using the Box- Behnken design to increase the production of xylooligosaccharides. The yield of xylooligosaccharides increased to 2.05- fold in the optimized medium than non-optimized medium. The produced xylooligosaccharides showed a peak at 890-900 cm-1 confirming the presence of β-glycosidic linkages when detected by Fourier Transform Infra-Red (FT IR). The prebiotic effect of produced xylooligosaccharides was studied on two strains named Lactococcus lactis Cl MN515342 and Lactococcus lactis Cp MN515343 isolated from curd samples. The growth of these Lactococcus strains was found to be stimulated. A log Colony Forming Unit/ml value of 7.813 and 8.146 for Cp and Cl was observed in a medium incorporated with XOS. The Short-Chain Fatty Acid (SCFA) production by Lactococcus lactis in a medium incorporated with xylooligosaccharides was detected by GCMS. The use of lignocellulosic waste for the production of xylooligosaccharides reduces the production cost. The xylooligosaccharides produced by Massilia timonae has shown promising result in stimulating the growth of Lactococcus lactis. Thus, XOS can be incorporated in curd or Yoghurt along with the Lactococcus lactis and used as synbiotic food.

Production of xylo-oligosaccharides (XOS) of tailored degree of polymerization from acetylated xylans through modelling of enzymatic hydrolysis

Xylo-oligosaccharides (XOS) are emerging prebiotics that have recently been gained a great interest in the market of functional foods. Since their beneficial activity strictly depends on their chemical structure and on their degree of polymerization (DP), in this work an enzymatic method was developed to produce XOS with variable and modellable DPs, involving a combination of a commercial endo-β-1,4-xylanase M3 from Trichoderma longibrachiatum and a deacetylase, using a commercial acetylated standard xylan as substrate. A Design of Experiment (DoE) was developed and through the variation of some hydrolysis conditions, some experiments allowed to obtain significant amounts of XOS with DP 7–10, up to 11%, despite XOS with DP 2–4 were always the most abundant (60–96% of total XOS). The most impacting parameter on the XOS distribution was the order of addition of the xylanase and deacetylating enzyme, while pH showed to have a great influence on the total yield. The method was also tested on an acetylated xylan extracted from grape stalks, structurally similar to the commercial standard xylan. The model was found to work in a very similar way also on the non-purified xylan sample, allowing the manipulation of enzymatic hydrolysis on a low-cost by-product, with the potential to obtain on a large scale XOS with high added value and with a specific DP, depending on the final application.