Oat Spelt Research Articles

Genome-wide transcriptomic analysis of a superior biomass-degrading strain of A. fumigatus revealed active lignocellulose-degrading genes.

BackgroundVarious saprotrophic microorganisms, especially filamentous fungi, can efficiently degrade lignocellulose that is one of the most abundant natural materials on earth. It consists of complex carbohydrates and aromatic polymers found in the plant cell wall and thus in plant debris. Aspergillus fumigatus Z5 was isolated from compost heaps and showed highly efficient plant biomass-degradation capability.ResultsThe 29-million base-pair genome of Z5 was sequenced and 9540 protein-coding genes were predicted and annotated. Genome analysis revealed an impressive array of genes encoding cellulases, hemicellulases and pectinases involved in lignocellulosic biomass degradation. Transcriptional responses of A. fumigatus Z5 induced by sucrose, oat spelt xylan, Avicel PH-101 and rice straw were compared. There were 444, 1711 and 1386 significantly differently expressed genes in xylan, cellulose and rice straw, respectively, when compared to sucrose as a control condition.ConclusionsCombined analysis of the genomic and transcriptomic data provides a comprehensive understanding of the responding mechanisms to the most abundant natural polysaccharides in A. fumigatus. This study provides a basis for further analysis of genes shown to be highly induced in the presence of polysaccharide substrates and also the information which could prove useful for biomass degradation and heterologous protein expression.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1658-2) contains supplementary material, which is available to authorized users.

Expression of Recombinant Bacillus amyloliquefaciens Xylanase A in Escherichia coli and Potential Application in Xylan Hydrolysis

Bacillus amyloliquefaciens xylanase A (baxA), an endoxylanase (EC. 3.2.1.8) gene, was cloned through PCR using the genome of B. amyloliquefaciens as a template. The open reading frame of baxA was 642 bp, and the gene encoded a 213 amino acid protein with a predicted molecular mass of 23.3 kDa. reBaxA1 produced in Escherichia coli with the pET30a(+) vector (T7 lac promoter) formed inclusion body and did not show any xylanase activity. reBaxA2 produced in E. coli with the pCold TF vector (cspA lac promoter) showed high xylanase activity; this enzyme was secreted into the culture medium and remained in the cell. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis analysis showed that the molecular weights of reBaxA1 and reBaxA2 were approximately 28.5 and 77.2 kDa, respectively. The optimal activity of reBaxA2 occurred at 55 °C and pH 6.0. Moreover, the Michaelis–Menten constant (Km) and maximal activity (Vmax) of reBaxA2 were 4.98 mg•ml-1 and 12.79 μmol•min–1•mL–1, respectively. High–performance liquid chromatography analysis showed that reBaxA2 released xylooligosaccharides from birchwood, beechwood, and oat spelt xylans, with xylopentaose, xylotriose, and xylotetraose as major products, respectively.

GH11 xylanase from Emericella nidulans with low sensitivity to inhibition by ethanol and lignocellulose-derived phenolic compounds.

An endo-β-1,4-xylanase (X22) was purified from crude extract of Emericella nidulans when cultivated on submerged fermentation using sugarcane bagasse as the carbon source. The purified protein was identified by mass spectrometry and was most active at pH and temperature intervals of 5.0-6.5 and 50-60°C, respectively. The enzyme showed half-lives of 40, 10 and 7 min at 28, 50 and 55°C, respectively, and pH 5.0. Apparent Km and Vmax values on soluble oat spelt xylan were 3.39 mg/mL and 230.8 IU/mg, respectively, while Kcat and Kcat/Km were 84.6 s(-1) and 25.0 s(-1) mg(-1) mL. Incubation with phenolic compounds showed that tannic acid and cinnamic acid had an inhibitory effect on X22 but no time-dependent deactivation. On the other hand, ferulic acid, 4-hydroxybenzoic acid, vanillin and p-coumaric acid did not show any inhibitory effect on X22 activity, although they changed X22 apparent kinetic parameters. Ethanol remarkably increased enzyme thermostability and apparent Vmax and Kcat values, even though the affinity and catalytic efficiency for xylan were lowered.

Characterization of a thermostable, specific GH10 xylanase from Caldicellulosiruptor bescii with high catalytic activity

Xylanase (EC 3.2.1.8) is one of the most important enzymes for the biodegradation of xylan. Since many industrial processes utilizing xylanase are operated at elevated temperatures, thermostable xylanases are highly desirable. In the present study, xyn10B gene from thermophilic bacterium Caldicellulosiruptor bescii that encodes a glycoside hydrolase (GH) family 10 xylanase was overexpressed in Escherichia coli and systematically characterized. CbXyn10B exhibited optimal activity at pH 7.2 and 70°C. It had a half-life of about 7.7h at 60°C, and retained over 85% of maximal activity after incubation at pH 4.0–12.0. The activity of this xylanase was not affected by most divalent cations, but inhibited by Fe3+ and Zn2+. CbXyn10B exhibited high activity on beech wood xylan, oat spelt xylan, and birch wood xylan, with specific activities of about 450Umg−1. Compared with other GH10 xylanases, CbXyn10B was highly specific for xylan and showed low catalytic efficiency toward sodium carboxymethyl cellulose and p-nitrophenyl-β-d-xylopyranoside. HPLC analysis of the products released from xylo-oligosaccharides and xylan revealed that xylobiose was the predominant hydrolytic product. The action mode of the enzyme was studied by product analysis, homology modeling and molecular docking to gain an insight into the structural basis for its substrate recognition mechanism.

Paenibacillus curdlanolyticus B-6 xylanase Xyn10C capable of producing a doubly arabinose-substituted xylose, α-l-Araf-(1 → 2)-[α-l-Araf-(1 → 3)]-d-Xylp, from rye arabinoxylan

Paenibacillus curdlanolyticus B-6 Xyn10C is a single module xylanase consisting of a glycoside hydrolase family-10 catalytic module. The recombinant enzyme, rXyn10C, was produced by Escherichia coli and characterized. rXyn10C was highly active toward soluble xylans derived from rye, birchwood, and oat spelt, and slightly active toward insoluble wheat arabinoxylan. It hydrolyzed xylooligosaccharides larger than xylotetraose to produce xylotriose, xylobiose, and xylose. When rye arabinoxylan and oat spelt xylan were treated with the enzyme and the hydrolysis products were analyzed by thin layer chromatography (TLC), two unknown hydrolysis products, U1 and U2, were detected in the upper position of xylose on a TLC plate. Electrospray ionization mass spectrometry and enzymatic analysis using Bacillus licheniformis α-l-arabinofuranosidase Axh43A indicated that U1 was α-l-Araf-(1→2)-[α-l-Araf-(1→3)]-d-Xylp and U2 was α-l-Araf-(1→2)-d-Xylp, suggesting that rXyn10C had strong activity toward a xylosidic linkage before and after a doubly arabinose-substituted xylose residue and was able to accommodate an α-1,2- and α-1,3-linked arabinose-substituted xylose unit in both the −1 and +1 subsites. A molecular docking study suggested that rXyn10C could accommodate a doubly arabinose-substituted xylose residue in its catalytic site, at subsite −1. This is the first report of a xylanase capable of producing α-l-Araf-(1→2)-[α-l-Araf-(1→3)]-d-Xylp from highly arabinosylated xylan.

Biological Activities of xylooligosaccharides generated from garlic straw xylan by purified xylanase from Bacillus mojavensis UEB-FK.

A newly isolated Bacterium strain named UEB-FK was selected from Tunisian Sahara, exhibiting the highest clear zone on agar plates containing oat spelt xylan by staining with Congo red. On the basis of 16S rDNA sequence analysis, this strain was identified as Bacillus mojavensis. This strain produced extracellular xylanase. Xylanase from the strain was purified to homogeneity and had an apparent molecular weight of 14 kDa. The K m and V max values of the purified xylanase on oat spelt xylan were 3.85 mg/mL and 250.02 U/mg, respectively. The optimum pH and temperature for the enzyme were found to be 4.0 and 50 °C, respectively, and the enzyme exhibited significant heat stability. In addition, the enzyme was found to be stable in a wide range of pH (3-9). The main hydrolysis products yielded from garlic straw-extracted xylan were xylobiose and xylotriose. The antioxidant and antibacterial activities of xylan oligosaccharide (XOS) were investigated. As regards to the in vitro antioxidant activities, the XOS showed a important DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activity (IC50 = 0.45 mg/mL) and a high β-carotene bleaching (IC50 = 2.2 mg/mL). Furthermore, XOS had a high antimicrobial activity against Klebsiella pneumoniae, Enterococcus faecalis, Bacillus thuringiensis, and Pseudomonas aeruginosa.

Biocatalytic properties and substrate-binding ability of a modular GH10 β-1,4-xylanase from an insect-symbiotic bacterium, Streptomyces mexicanus HY-14.

The gene (1350-bp) encoding a modular β-1,4-xylanase (XylU), which consists of an N-terminal catalytic GH10 domain and a C-terminal carbohydrate-binding module 2 (CBM 2), from Streptomyces mexicanus HY-14 was cloned and functionally characterized. The purified His-tagged recombinant enzyme (rXylU, 44.0 kDa) was capable of efficiently hydrolyze diverse xylosidic compounds, p-nitrophenyl-cellobioside, and p-nitrophenyl-xylopyranoside when incubated at pH 5.5 and 65°C. Especially, the specific activities (649.8 U/mg and 587.0 U/mg, respectively) of rXylU toward oat spelts xylan and beechwood xylan were relatively higher than those (<500.0 U/mg) of many other GH10 homologs toward the same substrates. The results of enzymatic degradation of birchwood xylan and xylooligosaccharides (xylotriose to xylohexaose) revealed that rXylU preferentially hydrolyzed the substrates to xylobiose (>75%) as the primary degradation product. Moreover, a small amount (4%<) of xylose was detected as the degradation product of the evaluated xylosidic substrates, indicating that rXylU was a peculiar GH10 β-1,4-xylanase with substrate specificity, which was different from its retaining homologs. A significant reduction of the binding ability of rXylU caused by deletion of the C-terminal CBM 2 to various insoluble substrates strongly suggested that the additional domain might considerably contribute to the enzyme-substrate interaction.

The characterization of the endoglucanase Cel12A from Gloeophyllum trabeum reveals an enzyme highly active on β-glucan.

The basidiomycete fungus Gloeophyllum trabeum causes a typical brown rot and is known to use reactive oxygen species in the degradation of cellulose. The extracellular Cel12A is one of the few endo-1,4-β-glucanase produced by G. trabeum. Here we cloned cel12A and heterologously expressed it in Aspergillus niger. The identity of the resulting recombinant protein was confirmed by mass spectrometry. We used the purified GtCel12A to determine its substrate specificity and basic biochemical properties. The G. trabeum Cel12A showed highest activity on β-glucan, followed by lichenan, carboxymethylcellulose, phosphoric acid swollen cellulose, microcrystalline cellulose, and filter paper. The optimal pH and temperature for enzymatic activity were, respectively, 4.5 and 50°C on β-glucan. Under these conditions specific activity was 239.2±9.1 U mg−1 and the half-life of the enzyme was 84.6±3.5 hours. Thermofluor studies revealed that the enzyme was most thermal stable at pH 3. Using β-glucan as a substrate, the Km was 3.2±0.5 mg mL−1 and the Vmax was 0.41±0.02 µmol min−1. Analysis of the effects of GtCel12A on oat spelt and filter paper by scanning electron microscopy revealed the morphological changes taking place during the process.

Behavior of cellulose and xylan in aqueous ammonia pretreatment.

The effect of aqueous ammonia on the solubilization of cellulose and xylans was investigated by detecting the amounts of reducing sugars and monosaccharides in the treatment liquors. The degree of cellulose and xylan solubilization increased with the increase of treatment temperature. When the treatment temperature increased from 20 to 90 °C, the amounts of reducing sugars released from Avicel and cellulose fiber by 21 % ammonia at a solid to liquid ratio of 1:10 for 24 h increased from 1.0 and 0.9 to 4.4 and 2.7 mg/g dry matter (DM), respectively. The amounts of reducing sugars released from wheat straw, beechwood, and oat spelt xylans increased from 1.2-7.0 to 3.3-13.5 mg/g DM. Xylans appeared to be more susceptible than cellulose in aqueous ammonia treatment. Structure analysis of untreated and treated Avicel and cellulose fiber showed that aqueous ammonia increased the specific surface area and crystallinity index of cellulose. Most of the cellulose and xylan that were solubilized existed in the form of oligomers such as cello-oligosaccharides and xylo-oligosaccharides. Xylobiose and xylotriose were the main oligosaccharides released from oat spelt xylan by aqueous ammonia treatment as confirmed by electrospray ionization-mass spectrometry. The results here indicated that a slight amount of cellulose and xylans was solubilized and low amounts of cellulase inhibitors, oligomers, were found during mild aqueous ammonia pretreatment process. Therefore, from the economical perspectives, mild ammonia pretreatment would be favorable for aqueous ammonia pretreatment of lignocelluloses.

Production of xylooligosaccharides from garlic straw xylan by purified xylanase from Bacillus mojavensis UEB-FK and their in vitro evaluation as prebiotics

A newly isolated Bacterium strain named UEB-FK was selected from Tunisian Sahara, exhibiting the highest clear zone on agar plates containing oat spelt xylan by staining with Congo red. On the basis of 16S rDNA sequence analysis, this strain was identified as Bacillus mojavensis. This strain produced extracellular xylanase. Xylanase from the strain was purified to homogeneity and had an apparent molecular weight of 14kDa. The Km and Vmax values of the purified xylanase on oat spelt xylan were 3.85mg/mL and 250.02U/mg, respectively. The optimum pH and temperature for the enzyme were found to be 4.0 and 50°C, respectively, and the enzyme exhibited significant heat stability even incubated in 60°C for 90min. In addition, the enzyme was found to be stable in a wide range of pH (3–9) and in presence of some detergents and organic solvents. The main hydrolysis products yielded from garlic straw extracted xylan were xylobiose and xylotriose. The resulting xylooligosaccharides (XOS) could be used as a special nutrient for lactic bacteria. Therefore, prebiotic effect of XOS was evaluated by in vitro fermentation using known probiotic strains viz. Bifidobacterium adolescentis and Lactobacillus acidophilus.

Novel alkali-tolerant GH10 endo-β-1,4-xylanase with broad substrate specificity from Microbacterium trichothecenolyticum HY-17, a gut bacterium of the mole cricket Gryllotalpa orientalis.

The XylH gene (1,167-bp) encoding a novel hemicellulase (41,584 Da) was identified from the genome of Microbacterium trichothecenolyticum HY-17, a gastrointestinal bacterium of Gryllotalpa orientalis. The enzyme consisted of a single catalytic domain, which is 74% identical to that of an endo-β-1,4-xylanase (GH10) from Isoptericola variabilis 225. Unlike other endo-β- 1,4-xylanases from invertebrate-symbiotic bacteria, rXylH was an alkali-tolerant multifunctional enzyme possessing endo-β-1,4-xylanase activity together with β-1,3/β-1,4- glucanase activity, which exhibited its highest xylanolytic activity at pH 9.0 and 60°C, and was relatively stable within a broad pH range of 5.0-10.0. The susceptibilities of different xylosebased polysaccharides to the XylH were assessed to be as follows: oat spelts xylan > beechwood xylan > birchwood xylan > wheat arabinoxylan. rXylH was also able to readily cleave p-nitrophenyl (pNP) cellobioside and pNP-xylopyranoside, but did not hydrolyze other pNP-sugar derivatives, xylobiose, or hexose-based materials. Enzymatic hydrolysis of birchwood xylan resulted in the product composition of xylobiose (71.2%) and xylotriose (28.8%) as end products.

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.

A new acidophilic endo-β-1,4-xylanase from Penicillium oxalicum: cloning, purification, and insights into the influence of metal ions on xylanase activity.

A new acidophilic xylanase (XYN11A) from Penicillium oxalicum GZ-2 has been purified, identified and characterized. Synchronized fluorescence spectroscopy was used for the first time to evaluate the influence of metal ions on xylanase activity. The purified enzyme was identified by MALDI TOF/TOF mass spectrometry, and its gene (xyn11A) was identified as an open reading frame of 706bp with a 68bp intron. This gene encodes a mature protein of 196 residues with a predicted molecular weight of 21.3kDa that has the 100% identity with the putative xylanase from the P. oxalicum 114-2. The enzyme shows a structure comprising a catalytic module family 10 (GH10) and no carbohydrate-binding module family. The specific activities were 150.2, 60.2, and 72.6 U/mg for beechwood xylan, birchwood xylan, and oat spelt xylan, respectively. XYN11A exhibited optimal activity at pH 4.0 and remarkable pH stability under extremely acidic condition (pH 3). The specific activity, K m and V max values were 150.2 U/mg, 30.7mg/mL, and 403.9μmol/min/mg for beechwood xylan, respectively. XYN11A is a endo-β-1,4-xylanase since it release xylobiose and xylotriose as the main products by hydrolyzing xylans. The activity of XYN11A was enhanced 155% by 1mM Fe(2+) ions, but was inhibited strongly by Fe(3+). The reason of enhancing the xylanase activity of XYN11A with 1mM Fe(2+) treatment may be responsible for the change of microenvironment of tryptophan residues studied by synchronous fluorescence spectrophotometry. Inhibition of the xylanase activity by Fe(3+) was first time demonstrated to associate tryptophan fluorescence quenching.

Purification and characterization of cellulase-free low molecular weight endo β-1,4 xylanase from an alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost.

Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is first report on actinomycetes that has the ability to produce thermostable cellulase-free xylanase, which is an important industrial enzyme used in the pulp and paper industry. Strain CKMX1 was characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis and 16Sr DNA and found to be C. cellulans CKMX1.The enzyme was purified by gel permeation and anion exchange chromatography and had a molecular mass of 29 kDa. Xylanase activity was optimum at pH 8.0 and 55 °C. The enzyme was somewhat thermostable, retaining 50 % of the original activity after incubation at 50 °C for 30 min. The xylanase had K m and V max values of 2.64 mg/ml and 2,000 µmol/min/mg protein in oat spelt xylan, respectively. All metal ions except HgCl2, CoCl2 as well as CdCl2 were well tolerated and did not adversely affect xylanase activity. The deduced internal amino acid sequence of C. cellulans CKMX1 xylanase by matrix assisted laser desorption ionization-time of flight mass spectrometry resembled the sequence of β-1,4-endoxylanase, which is a member of glycoside hydrolase family 11. Some of the novel characteristics that make this enzyme potentially effective in xylan biodegradation could be useful for pulp and paper biobleaching are discussed in this manuscript.

Manufacture of decyl pentosides surfactants by wood hemicelluloses transglycosidation: A potential pretreatment process for wood biomass valorization

We studied the reactivity of wood based hemicelluloses (beechwood xylan and poplar chips) in acid-catalyzed transglycosidation in decanol. Wood based hemicelluloses revealed to be more recalcitrant to transglycosidation than hemicelluloses from cereal origin (wheat straw, Oat spelt xylan). The chemical composition of the hemicelluloses, such as ash content or the level of arabinose and uronic acids in the polysaccharides structures, seems to influence the reactivity. The use of dimethylsulfoxide as solvent in the transglycosidation improved pentosides and glucosides yields for beechwood xylan, the more recalcitrant material. The surfactant properties of glycosides’ compositions produced from poplar were studied and compared to those of glycosides’ compositions obtained from pure monomeric carbohydrates or from wheat straw. The cellulose rich residue from poplar was hydrolyzed by concentrated acid saccharification yielding glucose in yields up to 55%.

Fusion of a xylan-binding module to gluco-oligosaccharide oxidase increases activity and promotes stable immobilization.

The xylan-binding module Clostridium thermocellum CBM22A was successfully fused to a gluco-oligosaccharide oxidase, GOOX-VN, from Sarocladium strictum via a short TP linker, allowing the fused protein to effectively bind different xylans. The presence of the CtCBM22A at the N-terminal of GOOX-VN increased catalytic activity on mono- and oligo-saccharides by 2-3 fold while not affecting binding affinity to these substrates. Notably, both GOOX-VN and its CBM fusion also showed oxidation of xylo-oligosaccharides with degrees of polymerization greater than six. Whereas fusion to CtCBM22A did not alter the thermostability of GOOX-VN or reduce substrate inhibition, CtCBM22A_GOOX-VN could be immobilized to insoluble oat spelt xylan while retaining wild-type activity. QCM-D analysis showed that the fused enzyme remained bound during oxidation. These features could be harnessed to generate hemicellulose-based biosensors that detect and quantify the presence of different oligosaccharides.

Purification and characterisation of thermo-alkaline pectinase enzyme from Hylocereus polyrhizus

The thermo-alkaline pectinase enzyme from Hylocereus polyrhizus was purified 232.3-fold with a 73.3 % recovery through ammonium sulphate precipitation, gel filtration, and ion exchange chromatography. Ion exchange chromatography combined with sodium dodecyl sulphate gel electrophoresis (SDS-PAGE) revealed that the enzyme was monomeric with a molecular weight of 34.2 kDa. The pectinase exhibited broad specificity towards polygalacturonic acid, arabinan, oat spelt xylan, and pNP-α-glucopyranoside. The optimum pH and temperature were 8.0 and 75 °C, respectively. This enzyme was stable over a wide pH range (3.0–11.0) and at relatively high temperature (85 °C for 1 h). The Km and Vmax values of pectinase towards polygalacturonic acid were 2.7 mg/ml and 34.30 U/mg proteins, respectively. In addition, the enzyme activity was inhibited by Ni2+, Al3+, and Fe2+ and was increased in the presence of Ca2+ and Mg2+ by 120 and 112 %, respectively. The purified pectinase demonstrated robust stability in response to surfactants and oxidising agents. EDTA, which is a powerful chelating agent, did not exert any significant effect on the enzyme stability. Thus, enzymes with these unique properties may be widely used in different types of industries and biotechnological applications.

Cloning, Expression, and Characterization of a Thermostable GH51 α-L-Arabinofuranosidase from Paenibacillus sp. DG-22

The gene encoding α-L-arabinofuranosidase (AFase) from Paenibacillus sp. DG-22 was cloned, sequenced, and expressed in Escherichia coli. The AFase gene (abfA) comprises a 1,509 bp open reading frame encoding 502 amino acids with a molecular mass of 56,520 daltons. The deduced amino acid sequence of the gene shows that AbfA is an enzyme consisting of only a catalytic domain, and that the enzyme has significant similarity to AFases classified into the family 51 of the glycosyl hydrolases. abfA was subcloned into the pQE60 expression vector to fuse it with a six-histidine tag and the recombinant AFase (rAbfA) was purified to homogeneity. The specific activity of the recombinant enzyme was 96.7 U/mg protein. Determination of the apparent molecular mass by gel-filtration chromatography indicated that AbfA has a tetrameric structure. The optimal pH and temperature of the enzyme were 6.0 and 60°C, respectively. The enzyme activity was completely inhibited by 1 mM HgCl2. rAbfA was active only towards p-nitrophephenyl α-L-arabinofuranoside and exhibited Km and Vmax values of 3.5 mM and 306.1 U/mg, respectively. rAbfA showed a synergistic effect in combination with endoxylanase on the degradation of oat spelt xylan and wheat arabinoxylan.

Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides Production

High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1 were detected when it was submerged-cultured. After three consecutive purification steps using Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase (XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according to SDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a 14.55-fold purification. The activity of XynNF was stimulated by Ca(2+), Ba(2+), DTT, and β-mercaptoethanol, but was inhibited by Fe(3+), Zn(2+), Fe(2+), Cu(2+), SDS, and EDTA. The purified XynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at 60°C and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at high temperature (70°C-80°C) and low pH range (pH 4.0-7.0), is potentially valuable for various industrial applications. The end products of high efficient oat spelt xylan hydrolysis by XynNF (an endoxylanase) containing 95.8% xylooligosaccharides of 2-4 degree of polymerization (DP2-4) with the enrichment of xylobiose (61.5%) indicated that XynNF is a promising candidate for xylooligosaccharides production.

Long-Term Physical Stability of Plasticized Hemicellulose Films

Oat spelt arabinoxylan (OsAX) and spruce galactoglucomannan (GGM) are hemicelluloses that can be extracted in large quantities from side-streams of the agriculture and forest industries. They both form self-standing films, making them potential future packaging materials. This systematic study focuses on the effect of long-term storage on the physical stability of hemicellulose-based films. OsAX and GGM films were plasticized with 40% (w/w of the polysaccharide) of glycerol, sorbitol, or their blends, and their stability was followed for four months. Ageing especially affected the glycerol-containing films, in which the tensile strength and Young’s modulus increased and elongation at break decreased. Although the mechanical properties were altered, storage did not affect crystallinity of the films. Oxygen gas permeability (OP) and water vapor permeability (WVP) properties were monitored in OsAX films. Interestingly WVP decreased during storage; more than a 40% decrease was seen when plasticizer blends contained 50% or more glycerol. In contrast, there were no drastic changes in the OP during storage; all the OPs obtained were between 3.7 and 8.9 [cm3 µm/ (m2 d kPa)].