Edible alginate-based films with anti-SARS-CoV-2 activity.

The viability of SARS-CoV-2 on food surfaces and its propagation through the food chain has been discussed by several stakeholders, as it may represent a serious public health problem, bringing new challenges to the food system. This work shows for the first time that edible films can be used against SARS-CoV-2. Sodium alginate-based films containing gallic acid, geraniol, and green tea extract were evaluated in terms of their antiviral activity against SARS-CoV-2. The results showed that all these films have strong in vitro antiviral activity against this virus. However, a higher concentration of the active compound (1.25%) is needed for the film containing gallic acid to achieve similar results to those obtained for lower concentrations of geraniol and green tea extract (0.313%). Furthermore, critical concentrations of the active compounds in the films were used to evaluate their stability during storage. Results showed that gallic acid-loaded films lose their activity from the second week of storage, while films with geraniol and green tea extract only show a drop in activity after four weeks. These results highlight the possibility of using edible films and coatings as antiviral materials on food surfaces or food contact materials, which may help to reduce the spreading of viruses through the food chain.

Inhibition of enzyme hydrolysis of cellulose by phenols from hydrothermally pretreated sugarcane straw.

Relatively few studies have addressed the characterization of sugarcane straw (SCS) for production of fermentable sugars through enzyme hydrolysis. Straw is a major co-product of the sugarcane harvest in Brazil that has potential to sustainably increase cellulosic feedstocks in Brazil by 50%. Pretreatment of 10% w/v straw with liquid hot water (LHW) at 180 °C for 50 min (severity, So, of 4.05), solubilizes hemicellulose, preserves glucan, and generates 4.49 g/L soluble phenolic compounds in the resulting liquid. Extracts from washing pretreated solids with excess hot water followed by acetone resulted in 1.10 and 0.83 g/L phenolics, respectively. Acetone-derived extracts were more inhibitory and decreased glucose yield for enzyme hydrolysis of Solka Floc (a lignin-free cellulose) by 42%. In comparison, pretreated straw washed with hot water or acetone was readily hydrolyzed to 92% and 97% by cellulase enzyme. Hydrothermally treated SCS has the potential to provide a valuable and added source of fermentable sugars suitable for bioprocessing into biofuels and bioproducts when cellulase enzyme inhibitors are removed after pretreatment.

Challenges in Using Ionic Liquids for Cellulosic Ethanol Production.

The growing need to expand the use of renewable energy sources in a sustainable manner, providing greater energy supply security and reducing the environmental impacts associated with fossil fuels, finds in the agricultural by-product bioethanol an economically viable alternative with significant expansion potential. In this regard, a dramatic boost in the efficiency of processes already in place is required, reducing costs, industrial waste, and our carbon footprint. Biofuels are one of the most promising alternatives to massively produce energy sustainably in a short-term period. Lignocellulosic biomass (LCB) is highly recalcitrant, and an effective pretreatment strategy should also minimize carbohydrate degradation by diminishing enzyme inhibitors and other products that are toxic to fermenting microorganisms. Ionic liquids (ILs) have been playing an important role in achieving cleaner processes as a result of their excellent physicochemical properties and outstanding performance in the dissolution and fractionation of lignocellulose. This review provides an analysis of recent advances in the production process of biofuels from LCB using ILs as pretreatment and highlighting techniques for optimizing and reducing process costs that should help to develop robust LCB conversion processes.

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.

Current Challenges for Biological Treatment of Pharmaceutical-Based Contaminants with Oxidoreductase Enzymes: Immobilization Processes, Real Aqueous Matrices and Hybrid Techniques.

The worldwide access to pharmaceuticals and their continuous release into the environment have raised a serious global concern. Pharmaceuticals remain active even at low concentrations, therefore their occurrence in waterbodies may lead to successive deterioration of water quality with adverse impacts on the ecosystem and human health. To address this challenge, there is currently an evolving trend toward the search for effective methods to ensure efficient purification of both drinking water and wastewater. Biocatalytic transformation of pharmaceuticals using oxidoreductase enzymes, such as peroxidase and laccase, is a promising environmentally friendly solution for water treatment, where fungal species have been used as preferred producers due to their ligninolytic enzymatic systems. Enzyme-catalyzed degradation can transform micropollutants into more bioavailable or even innocuous products. Enzyme immobilization on a carrier generally increases its stability and catalytic performance, allowing its reuse, being a promising approach to ensure applicability to an industrial scale process. Moreover, coupling biocatalytic processes to other treatment technologies have been revealed to be an effective approach to achieve the complete removal of pharmaceuticals. This review updates the state-of-the-art of the application of oxidoreductases enzymes, namely laccase, to degrade pharmaceuticals from spiked water and real wastewater. Moreover, the advances concerning the techniques used for enzyme immobilization, the operation in bioreactors, the use of redox mediators, the application of hybrid techniques, as well as the discussion of transformation mechanisms and ending toxicity, are addressed.

Challenges of Biomass Utilization for Bioenergy in a Climate Change Scenario.

The climate changes expected for the next decades will expose plants to increasing occurrences of combined abiotic stresses, including drought, higher temperatures, and elevated CO2 atmospheric concentrations. These abiotic stresses have significant consequences on photosynthesis and other plants' physiological processes and can lead to tolerance mechanisms that impact metabolism dynamics and limit plant productivity. Furthermore, due to the high carbohydrate content on the cell wall, plants represent a an essential source of lignocellulosic biomass for biofuels production. Thus, it is necessary to estimate their potential as feedstock for renewable energy production in future climate conditions since the synthesis of cell wall components seems to be affected by abiotic stresses. This review provides a brief overview of plant responses and the tolerance mechanisms applied in climate change scenarios that could impact its use as lignocellulosic biomass for bioenergy purposes. Important steps of biofuel production, which might influence the effects of climate change, besides biomass pretreatments and enzymatic biochemical conversions, are also discussed. We believe that this study may improve our understanding of the plant biological adaptations to combined abiotic stress and assist in the decision-making for selecting key agronomic crops that can be efficiently adapted to climate changes and applied in bioenergy production.

Ligninolytic enzymes production during polycyclic aromatic hydrocarbons degradation: effect of soil pH, soil amendments and fungal co-cultivation.

Soil microorganisms play an important role in the degradation of PAHs and use various metabolic pathways for this process. The effect of soil pH, different soil amendments and the co-cultivation of fungi on the degradation of PAHs in soil and on the activity of ligninolytic enzymes was evaluated. For that purpose, three fungi were studied: Trichoderma viride, Penicillium chrysogenum and Agrocybe aegerita. Biodegradation assays with a mixture of 200 ppm PAHs (fluorene, pyrene, chrysene, and benzo[a]pyrene-50 ppm each) were set up at room temperature for 8 weeks. The maximum laccase activity by solid state fermentation-SSF (7.43 U/g) was obtained by A. aegerita on kiwi peels with 2 weeks and the highest manganese peroxidase activity (7.21 U/g) was reached in 4 weeks, both at pH 7. Fluorene, pyrene, and benzo[a]pyrene achieved higher degradation rates in soil at pH 5, while chrysene was more degradable at pH 7. About 85-90% of the PAHs were degraded by fungal remediation. The highest degradation of fluorene was achieved by co-cultivation of A. aegerita and P. chrysogenum, remaining 14% undegradable. Around 13% of pyrene stay undegradable by A. aegerita and T. viride and by A. aegerita and P. chrysogenum, both systems supported in kiwi peels, while 11% of chrysene remained in soil by the co-cultivation of these fungi, supported by peanut shells. Regarding benzo[a]pyrene, 13% remained in soil after treatment with A. aegerita. Despite the increase in degradation of some PAHs with co-cultivation, higher enzyme production during degradation was observed when fungi were cultivated alone.

Development of a packed bed reactor for the removal of aromatic hydrocarbons from soil using laccase/mediator feeding system.

Polyaromatic hydrocarbons (PAH) are persistent pollutants of great concern due to their potential toxicity, mutagenicity and carcinogenicity. A biotechnological approach to remove PAH from soil was evaluated in this work using a laccase mediator system. Initially, laccase was produced by fungal co-cultivation, using kiwi peels as substrate. The produced laccase was applied to PAH contaminated soil to evaluate its efficiency on enzymatic bioremediation. Results showed that laccase mediator system was effective in the degradation of pyrene, fluorene, chrysene and a lower extension anthracene. Mediators improved the PAH degradation and natural mediators (ferulic acid and p-coumaric acid) were as effective as the synthetic mediator ABTS. However, the process was not effective in the benzo[a]pyrene degradation, one of the most recalcitrant and toxic PAH. This low degradation rate could be related to the low activity of the laccase mediator system in an environment lacking water. To overcome this issue, a PAH contaminated soil degradation system was developed in packed bed reactor (PBR) fed with laccase/mediator. Continuous flow of laccase/mediator improved the PAH degradation, achieving 74.8 %, 71.9 %, 72.2 %, 81.8 % and 100 % degradation for fluorene, anthracene, phenanthrene, chrysene and pyrene, respectively. This system was able to degrade 96 % benzo[a]pyrene, which was 90 % higher than the degradation in batch system. Results indicated that the produced laccase as well as the fed-batch degradation system developed in PBR could be successfully applied in the degradation of soil PAH pollutants, with the advantage of achieving higher degradation rates than in simple batch, as well as being a faster and simpler process than microorganism bioremediation.

Green synthesis of lignin nano- and micro-particles: Physicochemical characterization, bioactive properties and cytotoxicity assessment.

Lignin particles (LPs) have gained prominence due to their biodegradability and bioactive properties. LP production at nano and micro scale produced from organosolv lignin and the understanding of size's effect on their properties is unexplored. This work aimed to produce and characterize lignin nanoparticles and microparticles using a green synthesis process, based on ethanol-solubilized lignin and water. Spherical shape LPs, with a mean size of 75 nm and 215 nm and with a low polydispersity were produced, as confirmed by transmission electron microscopy and dynamic light scattering. LPs thermal stability improved over raw lignin, and the chemical structure of lignin was not affected by the production method. The antimicrobial tests proved that LPs presented a bacteriostatic effect on Escherichiacoli and Salmonella enterica. Regarding the antioxidant potential, LPs had a good antioxidant activity that increased with the reaction time and LPs concentration. LPs also presented an antioxidant effect against intracellular ROS, reducing the intracellular ROS levels significantly. Furthermore, the LPs showed a low cytotoxic effect in Caco-2 cell line. These results showed that LPs at different scales (nano and micro) present biological properties and are safe to be used in different high value industrial sectors, such as biomedical, pharmaceutical and food.

Nanocellulose Production: Exploring the Enzymatic Route and Residues of Pulp and Paper Industry.

Increasing environmental and sustainability concerns, caused by current population growth, has promoted a raising utilization of renewable bio-resources for the production of materials and energy. Recently, nanocellulose (NC) has been receiving great attention due to its many attractive features such as non-toxic nature, biocompatibility, and biodegradability, associated with its mechanical properties and those related to its nanoscale, emerging as a promising material in many sectors, namely packaging, regenerative medicine, and electronics, among others. Nanofibers and nanocrystals, derived from cellulose sources, have been mainly produced by mechanical and chemical treatments; however, the use of cellulases to obtain NC attracted much attention due to their environmentally friendly character. This review presents an overview of general concepts in NC production. Especial emphasis is given to enzymatic hydrolysis processes using cellulases and the utilization of pulp and paper industry residues. Integrated process for the production of NC and other high-value products through enzymatic hydrolysis is also approached. Major challenges found in this context are discussed along with its properties, potential application, and future perspectives of the use of enzymatic hydrolysis as a pretreatment in the scale-up of NC production.

Trametes versicolor laccase production using agricultural wastes: a comparative study in Erlenmeyer flasks, bioreactor and tray.

Laccases are very interesting biocatalysts of recognized importance for several industrial applications. Its production by Trametes versicolor, a white-rot fungus, was induced by a combination of cotton gin wastes (1%), a lignocellulosic waste, and vinasse (15%), an industrial by-product from sugarcane industry. The use of these agro-industrial wastes are interesting, since it helps in reducing the enzyme production costs, due to their low cost and wide availability, as well as the environmental contamination issues, due to their improper disposal. Thus, laccase production was studied in submerged fermentation of T. versicolor using these agro-industrial wastes (cotton gin waste and vinasse) as carbon source and an additional nitrogen source (0.1% peptone). Three different bioreactors were evaluated for laccase production, such as BioFlo 310 bioreactor, aluminium tray and Erlenmeyer flasks to achieve high levels of laccase production. The highest specific production of laccase was found in BioFlo 310 bioreactor with 12 days of fermentation (55.24 U/mg prot.), which has been shown to be closely related to the oxygen supply to the microorganism through aeration of the fermentation medium. This study brings new insights into green biotechnology regarding vinasse utilization, which is frequently discharged in soils, rivers, and lakes causing adverse effects on agricultural soils and biota, as well as the cotton gin waste recovery.

Enhancement and modeling of enzymatic hydrolysis on cellulose from agave bagasse hydrothermally pretreated in a horizontal bioreactor.

One of the major challenges in biofuels production from lignocellulosic biomass is the generation of high glucose titers from cellulose in the enzymatic hydrolysis stage of pretreated biomass to guarantee a cost-effective process. Therefore, the enzymatic saccharification on cellulose at high solid loading is an alternative. In this work, the agave bagasse was hydrothermally pretreated and optimized at 194 °C/30 min, obtaining a pretreated solid rich in cellulose content (>46.46%), and subjected to enzymatic hydrolysis at high solid levels. A horizontal bioreactor was designed for enzyme saccharification at high solid loadings [25% (w/v)]. The bioreactor improved mixing efficiency, with cellulose conversions up to 98% (195.6 g/L at 72 h). Moreover, mathematical modeling of cellulase deactivation demonstrated that cellulases lose most of their initial activity in the first hours of the reaction. Also, cellulose was characterized by X-ray diffraction, and the pretreated solids were visualized using scanning electron microscopy.

Lignin from an integrated process consisting of liquid hot water and ethanol organosolv: Physicochemical and antioxidant properties.

Corncob was successively pretreated by liquid hot water (LHW) and ethanol organosolv (EO) in an integrated process. LHW was performed at 200 °C for 30 min, and EO was performed using uncatalyzed ethanol-water solutions, according to a design of experiments. The effects of the most influential operational variables (ethanol concentration, temperature and time) on yield and chemical composition of the fractions were assessed. Results showed the factor with the greatest effect was ethanol concentration (p < 0.05), leading to a high-purity lignin (86.7%-93.1%) with a total phenolic content of around 25 mg GAE/g. Moreover, the solids recovered from the high ethanol concentration conditions presented the lowest lignin contents (15.4%-17.2%) with good preservation of cellulose (82.5%-88.6% of glucans). The lignin antioxidant capacity showed that all lignin samples presented radical scavenging activity (IC50 of 0.17 mg/mL and 0.016 mg/mL on DPPH (2,2­diphenyl­1­picrylhydrazyl) and ABTS (2,2'­azino­bis(3­ethylbenzothiazoline­6­sulphonic acid) assays, respectively) with values close to the commercial antioxidant BHT. Moreover, the chemical (FTIR) and thermal (DSC and TGA) characterization showed lignins with similar properties that were compared with lignin from direct ethanol organosolv process. Results showed that the integrated process of LHW-EO was the most effective way to obtain lignin with high purity and antioxidant capacity.

Cellulose nanocrystals from grape pomace: Production, properties and cytotoxicity assessment.

Cellulose nanocrystals (CNCs) were obtained from grape pomace through chemical and physical pretreatments. Bleached cellulose pulp was subjected to acid hydrolysis (AH) for 30 or 60 min and an ultrasound treatment to obtain CNCs (AH30S and AH60S). Compositional analyses of untreated (UGP) and pretreated (PGP) grape pomace showed the effectiveness of pretreatment in removing non-cellulosic components, recovering 80.1% cellulose in PGP (compared to 19.3% of UGP). Scanning and transmission electron microscopies were used to evaluate the CNCs morphology. AH in combination with ultrasound treatment led to needle-shaped structures and apparently more dispersed suspensions. Crystallinity index and thermal stability were studied by X-ray diffraction and thermogravimetric analysis, respectively. The AH60S sample presented high aspect ratio, crystallinity and thermal stability. CNCs toxicity was evaluated by exposing Caco-2 cells to CNCs suspension and evaluating their viability. Results showed that CNCs are non-toxic, opening the opportunity for their use on food and pharmaceutical applications.

Multi-step approach to add value to corncob: Production of biomass-degrading enzymes, lignin and fermentable sugars.

This work presents an integrated and multi-step approach for the recovery and/or application of the lignocellulosic fractions from corncob in the production of high value added compounds as xylo-oligosaccharides, enzymes, fermentable sugars, and lignin in terms of biorefinery concept. For that, liquid hot water followed by enzymatic hydrolysis were used. Liquid hot water was performed using different residence times (10-50min) and holding temperature (180-200°C), corresponding to severities (log(R0)) of 3.36-4.64. The most severe conditions showed higher xylo-oligosaccharides extraction (maximum of 93%) into the hydrolysates and higher recovery of cellulose on pretreated solids (maximum of 65%). Subsequently, hydrolysates and solids were used in the production of xylanases and cellulases, respectively, as well as, pretreated solids were also subjected to enzymatic hydrolysis for the recovery of lignin and fermentable sugars from cellulose. Maximum glucose yield (100%) was achieved for solids pretreated at log(R0) of 4.42 and 5% solid loading.

Liquid hot water pretreatment of multi feedstocks and enzymatic hydrolysis of solids obtained thereof.

Agricultural feedstocks (brewers' spent grain - BSG, corncob - CC, corn husk - CH, wheat straw - WS and Luffa sponge - LS) were pretreated by liquid hot water (LHW) in order to increase cellulose recovery and enzymatic saccharification. LHW-pretreatment resulted in hemicellulose solubilization, and solids enriched in cellulose. Chemical analysis showed different susceptibilities of the feedstocks to LHW-pretreatment and enzymatic hydrolysis. Pretreated feedstocks presented higher crystallinity (determined through X-ray diffraction) and thermal stability (determined through thermogravimetric analysis) than untreated feedstocks. SEM images confirmed the effect of LHW-pretreatment on structural changes. Moreover, enzymatic hydrolysis and cellulose conversion to glucose (CCG) were improved for pretreated feedstocks, with exception of LS. CCG (in relation to glucose potential on solids) followed the order: BSG>CH>WS>CC>LS. LHW-pretreatment showed to be a good technology to pretreat multi feedstocks and for improving the enzymatic hydrolysis of recalcitrant agricultural feedstocks to sugars, which can be further converted to ethanol-fuel and other value-added chemicals.

Effect of phenolic compounds from pretreated sugarcane bagasse on cellulolytic and hemicellulolytic activities.

This work shows both cellulases and hemicellulases are inhibited and deactivated by water-soluble and acetone extracted phenolics from sugarcane bagasse pretreated at 10% (w/v) for 30 min in liquid hot water at 180 or 200°C. The dissolved phenolics in vacuum filtrate increased from 1.4 to 2.4 g/L as temperature increased from 180 to 200°C. The suppression of cellulose and hemicellulose hydrolysis by phenolics is dominated by deactivation of the ß-glucosidase or ß-xylosidase components of cellulase and hemicellulase enzyme by acetone extract at 0.2-0.65 mg phenolics/mg enzyme protein and deactivation of cellulases and hemicellulases by the water soluble components in vacuum filtrate at 0.05-2mg/mg. Inhibition was a function of the type of enzyme and the manner in which the phenolics were extracted from the bagasse.

Purification and biochemical properties of multiple xylanases from Aspergillus ochraceus tolerant to Hg2+ ion and a wide range of pH.

Production of multiple xylanases, in which each enzyme has a specific characteristic, can be one strategy to achieve the effective hydrolysis of xylan. Three xylanases (xyl 1, xyl 2, and xyl 3) from Aspergillus ochraceus were purified by chromatography using diethylaminoethyl (DEAE) cellulose, Biogel P-60, and Sephadex G-100 columns. These enzymes are glycoproteins of low molecular weight with an optimum temperature at 60 °C. The glycosylation presented is apparently not related to thermostability, since xyl 3 (20 % carbohydrate) was more thermostable than xyl 2 (67 % carbohydrate). Xyl 3 was able to retain most of its activity in a wide range of pH (3.5-8.0), while xyl 1 and xyl 2 presented optimum pH of 6.0. Xyl 1 and xyl 2 were activated by 5 and 10 mM MnCl2 and CoCl2, while xyl 3 was activated by 1 mM of the same compounds. Interestingly, xyl 2 presented high tolerance toward mercury ion. Xylanases from A. ochraceus hydrolyzed xylans of different origins, such as birchwood, oat spelt, larchwood, and eucalyptus (around 90 % or more), except xyl 2 and xyl 3 that hydrolyzed with lesser efficiency eucalyptus (66.7 %) and oat spelt (44.8 %) xylans.

Purification, partial characterization, and covalent immobilization-stabilization of an extracellular α-amylase from Aspergillus niveus.

An extracellular amylase secreted by Aspergillus niveus was purified using DEAE fractogel ion exchange chromatography and Sephacryl S-200 gel filtration. The purified protein migrated as a single band in 5 % polyacrylamide gel electrophoresis (PAGE) and 10 % sodium dodecyl sulfate (SDS-PAGE). The enzyme exhibited 4.5 % carbohydrate content, 6.6 isoelectric point, and 60 and 52 kDa molar mass estimated by SDS-PAGE and Bio-Sil-Sec-400 gel filtration column, respectively. The amylase efficiently hydrolyzed glycogen, amylose, and amylopectin. The end-products formed after 24 h of starch hydrolysis, analyzed by thin layer chromatography, were maltose, maltotriose, maltotetraose, and maltopentaose, which classified the studied amylase as an α-amylase. Thermal stability of the α-amylase was improved by covalent immobilization on glyoxyl agarose (half-life of 169 min, at 70 °C). On the other hand, the free α-amylase showed a half-life of 20 min at the same temperature. The optima of pH and temperature were 6.0 and 65 °C for both free and immobilized forms.

A novel xylan degrading ß-D-xylosidase: purification and biochemical characterization.

Aspergillus ochraceus, a thermotolerant fungus isolated in Brazil from decomposing materials, produced an extracellular ß-xylosidase that was purified using DEAE-cellulose ion exchange chromatography, Sephadex G-100 and Biogel P-60 gel filtration. ß-xylosidase is a glycoprotein (39 % carbohydrate content) and has a molecular mass of 137 kDa by SDS-PAGE, with optimal temperature and pH at 70 °C and 3.0-5.5, respectively. ß-xylosidase was stable in acidic pH (3.0-6.0) and 70 °C for 1 h. The enzyme was activated by 5 mM MnCl2 (28 %) and MgCl2 (20 %) salts. The ß-xylosidase produced by A. ochraceus preferentially hydrolyzed p-nitrophenyl-ß-D-xylopyranoside, exhibiting apparent K(m) and V(max) values of 0.66 mM and 39 U (mg protein)⁻¹ respectively, and to a lesser extent p-nitrophenyl-ß-D-glucopyranoside. The enzyme was able to hydrolyze xylan from different sources, suggesting a novel ß-D-xylosidase that degrades xylan. HPLC analysis revealed xylans of different compositions which allowed explaining the differences in specificity observed by ß-xylosidase. TLC confirmed the capacity of the enzyme in hydrolyzing xylan and larger xylo-oligosaccharides, as xylopentaose.