The DUF579 proteins GhIRX15s regulate cotton fiber development by interacting with proteins involved in xylan synthesis

Cotton provides the most abundant natural fiber for the textile industry.The mature cotton fiber largely consists of secondary cell walls with the highest proportion of cellulose and a small amount of hemicellulose and lignin.To dissect the roles of hemicellulosic polysaccharides during fiber development,four IRREGULAR XYLEM 15(IRX15)genes,GhIRX15-1/-2/-3/-4,were functionally characterized in cotton.These genes encode DUF579 domain-containing proteins,which are homologs of AtIRX15 involved in xylan biosynthesis.The four GhIRX15 genes were predominantly expressed during fiber secondary wall thickening,and the encoded proteins were localized to the Golgi apparatus.Each GhIRX15 gene could restore the xylan deficient phenotype in the Arabidopsis irx15irx15l double mutant.Silencing of GhIRX15s in cotton resulted in shorter mature fibers with a thinner cell wall and reduced cellulose content as compared to the wild type.Intriguingly,GhIRX15-2 and GhIRX15-4 formed homodimers and heterodimers.In addition,the GhIRX15s showed physical interaction with glycosyltransferases GhGT43C,GhGT47A and GhGT47B,which are responsible for synthesis of the xylan backbone and reducing end sequence.Moreover,the GhIRX15s can form heterocomplexes with enzymes involved in xylan modification and side chain synthesis,such as GhGUX1/2,GhGXM1/2 and GhTBL1.These findings suggest that GhIRX15s participate in fiber xylan biosynthesis and modulate fiber development via forming large multiprotein complexes.

Study on the green extraction of corncob xylan by deep eutectic solvent

Corn as one of the world's major food crops,its by-product corn cob is also rich in resources.However,the unreasonable utilization of corn cob often causes the environmental pollution,waste of resources and other problems.As one of the most abundant polymers in nature,xylan is widely used in food,medicine,materials and other fields.Corn cob is rich in xylan,which is an ideal raw material for extracting xylan.However,the intractable lignin is covalently linked to xylan,which increases the difficulty of xylan extraction.It has been reported that the deep eutectic solvent(DES)could preferentially dissolve lignin in biomass,thereby dissolving the xylan.Then,the xylan in the extract was separated by ethanol precipitation method.The xylan precipitate was obtained after centrifugation,while the supernatant was retained.The components of the supernatant after ethanol precipitation were separated by the rotary evaporator.The ethanol,water and DES were collected for the subsequent extraction of corn cob xylan.In this study,a novel way was provided for the green production of corn cob xylan.The DES was used to extract xylan from corn cob which was used as the raw material.The effects of solid-liquid ratio,reaction time,reaction temperature and water content of DES on the extraction rate of corn cob xylan were investigated by the single factor test.Furthermore,the orthogonal test was designed to optimize the xylan extraction process.The structure of corn cob xylan was analyzed and verified.The results showed that the optimum extraction conditions of corn cob xylan were as follows:the ratio of corn cob to DES was 1:15(g:mL),the extraction time was 3 h,the extraction temperature was 60℃,and the water content of DES was 70%.Under these conditions,the extraction rate of xylan was 16.46%.The extracted corn cob xylan was distinctive triple helix of polysaccharide,which was similar to the structure of commercially available xylan.Xylan was effectively and workably extracted from corn cob by the DES method.This study provided a new approach for high value conversion of corn cob and the clean production of xylan.

Paddy Husk as Support for Solid State Fermentation to Produce Xylanase from Bacillus pumilus

To optimize culture conditions for xylanase production by solid state fermentation （SSF） using Bacillus pumilus, with paddy husk as support, solid medium contained 200 g of paddy husk with 800 mL of liquid fermentation medium [xylan, 20.0 g/L; peptone, 2.0 g/L; yeast extract, 2.5 g/L; K2HPO4, 2.5 g/L; KH2PO4, 1.0 g/L; NaCl, 0.1 g/L; （NH4）2SO4, 2.0 g/L, CaCl2-2H2O, 0.005 g/L; MgCl2.6H2O, 0.005 g/L; and FeCI3, 0.005 g/L] at pH 9.0 was applied. The highest xylanase activity （142.0 ±0.47 U/g DM] was obtained on the 6th day at 30℃ The optimized paddy husk to liquid fermentation medium ratio was 2：9, and the optimized culture temperature was 40℃. When commercial Birchwood xylan was replaced with different concentrations of corncob, xylanase production was maximized （224.2 U/g DM） in the medium with 150 g/L corncob. Xylanase production was increased by sucrose, fructose and arabinose, whereas reduced by glucose, galactose, lactose and amylose. When organic nitrogen sources were replaced with locally available nitrogen sources such as groundnut powder or sesame seedcake powder or coconut seedcake powder or soy meal powder, the highest xylanase production （290.7 U/g DM） was obtained in the medium with soy meal powder and 16.0 g/L of soy meal powder was the optimum （326.5±0.34 U/g DM）. Based on the optimization studies, B. pumilus produced 2.3 times higher xylanase activity. The medium cost was reduced from 2 458.3 to 178.3 SLR/kg and the total activity which could be obtained from 1 kg of the medium was increased from 48 624 to 220 253 Units.

Effects of protein and lignin on cellulose and xylan anaylses of lignocellulosic biomass

Interactions of lignocellulosic components during fiber analysis were investigated using the highly adopted compositional analysis procedure from the National Renewable Energy Laboratory（NREL）,USA.Synthetic feedstock samples were used to study the effects of lignin/protein,cellulose/protein,and xylan/protein interaction on carbohydrate analysis.Disregarding structural influence in the synthetic samples,lignin and protein components were the most significant（P〈0.05）factors on cellulose analysis.Measured xylan was consistent and unaffected by content variation throughout the synthetic analysis.Validation of the observed relationships from synthetic feedstocks was fulfilled using real lignocellulosic feedstocks：corn stover,poplar,and alfalfa,in which similar results have been obtained,excluding cellulose analysis of poplar under higher protein content and xylan analysis of alfalfa under higher protein content.The results elucidated that according to their protein and lignin contents of different lignocellulosic materials,accuracy of the NREL method on cellulose and xylan analyses could be improved by applying a stronger extraction step to replace water/ethanol extraction.

High Oxygen Nanocomposite Barrier Films Based on Xylan and Nanocrystalline Cellulose

The goal of this work is to produce nanocomposite film with low oxygen permeability by casting an aqueous solution containing xylan,sorbitol and nanocrystalline cellulose.The morphology of the resulting nanocomposite films was examined by scanning electron microscopy and atomic force microscopy which showed that control films containing xylan and sorbitol had a more open structure as compared to xylan-sorbitol films containing sulfonated nanocrystalline cellulose.The average pore diameter,bulk density,porosity and tortuosity factor measurements of control xylan films and nanocomposite xylan films were examined by mercury intrusion porosimetry techniques.Xylan films reinforced with nanocrystalline cellulose were denser and exhibited higher tortuosity factor than the control xylan films.Control xylan films had average pore diameter,bulk density,porosity and tortuosity factor of 0.1730 μm,0.6165 g/ml,53.0161% and 1.258,respectively as compared to xylan films reinforced with 50% nanocrystalline cellulose with average pore diameter of 0.0581 μm,bulk density of 1.1513 g/ml,porosity of 22.8906% and tortuosity factor of 2.005.Oxygen transmission rate tests demonstrated that films prepared with xylan,sorbitol and 5%,10%,25% and 50% sulfonated nanocrystalline cellulose exhibited a significantly reduced oxygen permeability of 1.1387,1.0933,0.8986 and 0.1799 cm^3×μm/m^2×d×k Pa respectively with respect to films prepared solely from xylan and sorbitol with a oxygen permeability of 189.1665 cm^3×μm/m^2×d×k Pa.These properties suggested these nanocomposite films have promising barrier properties.

Global identification of genes associated with xylan biosynthesis in cotton fiber

Background:Mature cotton fiber secondary cell wall comprises largely of cellulose(>90%)and small amounts of xylan and lignin.Little is known about the cotton fiber xylan biosynthesis by far.Results:To comprehensively survey xylan biosynthetic genes in cotton fiber,we identified five IRX9,five IRX10,one IRX14,six IRX15,two FRA8,one PARVUS,eight GUX,four GXM,two RWA,two AXY9,13 TBL genes by using phylogenetic analysis coupled with expression profile analysis and co-expression analyses.In addition,we also identified two GT61 members,two GT47 members,and two DUF579 family members whose homologs in Arabidopsis were not functionally characterized.These 55 genes were regarded as the most probable genes to be involved in fiber xylan biosynthesis.Further complementation analysis indicated that one IRX10 like and two FRA8 related genes were able to partially recover the irregular xylem phenotype conferred by the xylan deficiency in their respective Arabidopsis mutant.We conclude that these genes are functional orthologs of respective genes that are implicated in GX biosynthesis.Conclusion:The list of 55 cotton genes presented here provides not only a solid basis to uncover the biosynthesis of xylan in cotton fiber,but also a genetic resource potentially useful for future studies aiming at fiber improvement via biotechnological approaches.

Facile Approach for Preparation of Xylan-based Double-network Hydrogels

In this study,xylan-based double-network(DN)hydrogels(xylanbased DN gels)with excellent mechanical properties were prepared using acrylic acid and acrylamide(AM)based on a DN approach.The first layer network was obtained by grafting and crosslinking polyacrylic acid(PAA)molecular chains onto xylan with ammonium persulfate(APS)as the initiator and N,N'-methylenebisacrylamide(MBA)as the crosslinking agent;this network was subsequently immersed into an aqueous AM monomer in the presence of APS and MBA for the preparation of the second layer network.The results showed that the double networks were crosslinked by covalent bonds and that the mechanical properties of the xylan-based DN gels were enhanced.Thus,the xylan-based DN gels exhibited a maximum compression stress of 24.9 MPa.The xylan-based DN gels could also recover 97%of their original height after 15 repeated compression cycles;this indicates that the xylan-based DN gels possessed high resistance to friction and wear.Therefore,the prepared xylan-based DN gels have considerable potential for tissue engineering applications.

Optimization of Microwave Assisted Alkaline Extraction of Xylan from Birch Wood Using Response Surface Methodology

The main purpose of this study was to optimize microwave assisted alkaline extraction of the hemicellulose, xylan, from birch wood. The simultaneous effects of process variables such as time (10 - 30 minutes), concentration of sodium hydroxide solution (4 - 8 wt%), solid to liquid ratio (1:8 to 1:20, g:mL), and sample size (5 - 10 g) on the temperature of the wood slurry, wood dissolution, and yield of extraction were evaluated. A central composite design (CCD) and response surface methodology (RSM) were used for the optimization of the extraction process. Based on the CCD, quadratic models were developed to correlate the extraction process variables with the responses such as temperature of wood slurry, wood dissolution, and yield of xylan and the models were analyzed using appropriate statistical methods (ANOVA). Statistical analysis showed that all the models developed were found to be adequate for the prediction of the respective responses. Optimization of the process was performed using a numerical optimization available in the software to maximize the yield of xylan and the optimum process variables for the maximum yield of xylan was found to be: 10 g of wood fibres, 8 wt% of NaOH solution, 1:10 solid to liquid ratio (g:mL) and 25 minutes of irradiation time. About 72.5% of the xylan present in the birch wood was extracted using the optimized extraction parameters.

Xylanase-producing microflora in Eastern Ghats of Andhra Pradesh,India

We screened soil samples collected from underneath shrubs and/or large trees at different locations in the Eastern Ghats of Andhra Pradesh for xylanase-producing microorganisms. Xylose-utilizing bacteria were numerically dominant in soils of most locations whereas xylose-and xylan-utilizing actinobacteria were minor components. Xylan-utilizing fungi constituted a major share of total microbial populations in soil samples collected at half of the sites, whereas xylan-utilizing bacteria were predominant at other sampling locations. Some of the isolates of fungi exhibited xylanase activity with a range of400–4000 U/ml, indicating great potential for their uses in paper, pulping and bioethanol industries for producing value-added products.

Structural Characteristics of Cellulose and Xylan during in vitro Fermentation by Pig Fecal Bacteria

In this study,cellulose and xylan were in vitro fermented by pig fecal bacteria.Rapid fermentation(40 h) and extended fermentation(eight weeks)were performed.The properties and ultra-structure changes of post-fermented solid residues were studied.In the end effluent,acetic acid,propionic acid,and butyric acid were observed to be the principal short-chain fatty acids(SCFAs) produced by anaerobic fermentation.Xylan was more accessible to bacteria than cellulose,leading to higher SCFA and lactic acid production.In addition,the crystalline structure of cellulose changed,leading to 16.3% and42.1% increases in crystallinity index for rapid and extended fermentation,respectively.Through this research,a systematic and advanced method to study the degradation chemistry of cellulose and xylan during fermentation was developed.

一种Xylan表面喷涂的工艺规程及其检验方法

金属表面处理技术可以在不增加或增加较少成本的情况下,使工件表面达到被保护和强化的效果,从而提高产品的使用寿命和稳定性,改善产品的使用性能和质量,增强产品的竞争能力,石油机械行业中金属组件和紧固件的表面处理方式多种多样,总体来说主要通过表面涂层技术在基体表面制备各种镀、涂覆层以及通过各种表面改性技术改变基体表面的组织和性能的两种方式,但都因工艺过程繁杂、化学反应效果不可控等原因存在着一些问题,如电镀方式处理不当可能导致工件发生氢脆现象或造成污染环境等后果。

Xylan公司首次将IP防火墙功能集成于校园交换网

美国Xylan公司近日宣布，首次为校园交换网提供IP防火墙功能。这种新功能将被集成于Xylan的Om-ni-Switch系列和Pizza-Switch系列。

Molecular Dissection of Xylan Biosynthesis during Wood Formation in Poplar

Xylan, being the second most abundant polysaccharide in dicot wood, is considered to be one of the factors contributing to wood biomass recalcitrance for biofuel production. To better utilize wood as biofuel feedstock, it is crucial to functionally characterize all the genes involved in xylan biosynthesis during wood formation. In this report, we investigated roles of poplar families GT43 and GT8 glycosyltransferases in xylan biosynthesis during wood formation. There exist seven GT43 genes in the genome of poplar （Populus trichocarpa）, five of which, namely PtrGT43A, PtrGT43B, PtrGT43C, PtrGT43D, and PtrGT43E, were shown to be highly expressed in the developing wood and their encoded proteins were localized in the Golgi. Comprehensive genetic complementation coupled with chemical analyses demonstrated that overexpression of PtrGT43A/B/E but not PtrGT43C/D was able to rescue the xylan defects conferred by the Arabidopsis irx9 mutant, whereas overexpression of PtrGT43C/D but not PtrGT43A/B/E led to a complementation of the xylan defects in the Arabidopsis irx14 mutant. The essential roles of poplar GT43 members in xylan biosynthesis was further substantiated by RNAi down-regulation of GT43B in the hybrid poplar （Populus alba x tremula） leading to reductions in wall thickness and xylan content in wood, and an elevation in the abundance of the xylan reducing end sequence. Wood digestibility analysis revealed that cellulase digestion released more glucose from the wood of poplar GT43B RNAi lines than the control wood, indicating a decrease in wood biomass recalcitrance. Furthermore, RNAi down-regulation of another poplar wood-associated glycosyltransferase, PoGTSD, was shown to cause decreases in wall thickness and xylan content as well as in the abundance of the xylan reducing end sequence. Together, these findings demonstrate that the poplar GT43 members form two functionally non-redundant groups, namely PtrGT43A/B/E as functional orthologs of Arabidopsis IRX9 and PtrGT43C/D as functional orthologs ofArabidopsis IRX14, all of which are involved in the biosynthesis of xylan backbones, and that the poplar GT8D is essential for the biosynthesis of the xylan reducing end sequence.

IRX14 and IRX14-LIKE, Two Glycosyl Transferases Involved in Glucuronoxylan Biosynthesis and Drought Tolerance in Arabidopsis

IRX14 and IRX14-LIKE （IRX14L） are two closely related glycosyl transferases in the glycosyl transferase 43 （GT43） family of Arabidopsis. A T-DNA insertion mutant for IRX14 results in comparatively minor changes, such as irregular xylem, while a mutation for IRX14L results in no changes. However, an irx14 and irx14L double mutant severely affects growth and development, with the dwarf plants failing to produce an inflorescence stem. Plants that are homozygous for IRX14 but heterozygous for IRX14L （irx14 irx14L（±）） exhibit an intermediate phenotype, including noticeably smaller leaves, stems, and underdeveloped siliques. Additionally, the T-DNA insertion mutant for IRX14 was found to result in a drought-tolerant phenotype. Carbohydrate analysis of total cell wall extracts revealed a reduction in xylose for the irx14 and irx14 irx14L（±） mutants, consistent with a defect in glucuronoxylan biosynthesis. Immunolocalization of xylan with the LM10 antibody revealed a loss of xylan in irx14 mutants and a further reduction in the irx14 irx14L（±） mutants. IRX14L likely functions redundantly with IRX14 in glucuronoxylan biosynthesis, with IRX14 having a more important role in the process.

Mechanism of Xylan Pyrolysis by Py-GC/MS

In order to investigate the decomposition behavior of hemicellulose, xylan was chosen as the representative of hemicellulose to study the fast pyrolysis on the combination system of analytical pyrolyzer and gas chromatograph coupled with mass spectrometer（Py-GC/MS）. The main condensable products of xylan pyrolysis consisted of acids, al- dehydes, and ketones; while gas products contained CO2, CO, CH4 and H〉 Acetic acid and furfural were the most abun- dant products with the highest contents of 20.11% and 20.24% respectively. While furfural and acetic acid were formed competitively with residence time and temperature increases, the distribution of xylan pyrolysis products did not vary with the residence time and temperature, while the total content of several kinds of products changed a lot. According to the analysis of experimental data, a reaction pathway of xylan decomposition was deduced so as to illustrate the forma- tion mechanism of main products.

Conversion of xylose and xylan into furfural in biorenewable choline chloride–oxalic acid deep eutectic solvent with the addition of metal chloride

An environmentally benign processing approach for furfural production from xylose and xylan under very mild conditions（353–373 K） was developed with the addition of metal chlorides in ChCl–oxalic acid（a deep eutectic solvent（DES）） synthesized from cheap and renewable starting materials）. ChCl–oxalic acid acted as both a Br？nsted acid catalyst and a reaction medium in this catalytic route. In addition, a biphasic system with methyl isobutyl ketone as an extracting reagent（DES/MIBK） to further increase furfural yield was also proposed. This processing approach for producing furfural eliminated the large energy consumption for high pressure saturated steam and the generation of acidic effluent, which was very difficult to handle. The whole catalytic system was more environmentally friendly compared with the commercial process for furfural production.

Role of UDP-Glucuronic Acid Decarboxylase in Xylan Biosynthesis in Arabidopsis

UDP-xylose （UDP-Xyl） is the Xyl donor used in the synthesis of major plant cell-wall polysaccharides such as xylan （as a backbone-chain monosaccharide） and xyloglucan （as a branching monosaccharide）. The biosynthesis of UDP-Xyl from UDP-glucuronic acid （UDP-GIcA） is irreversibly catalyzed by UDP- glucuronic acid decarboxylase （UXS）. Until now, little has been known about the physiological roles of UXS in plants. Here, we report that AtUXS1, AtUXS2, and AtUXS4 are located in the Golgi apparatus whereas AtUXS3, AtUXS5, and AtUXS6 are located in the cytosoh Although all six single AtUXS T-DNA mu- tants and the uxsl usx2 uxs4 triple mutant show no obvious phenotype, the uxs3 uxs5 uxs6 triple mutant has an irregular xylem phenotype. Monosaccharide analysis showed that Xyl levels decreased in uxs3 uxs5 uxs6 and linkage analysis confirmed that the xylan content in uxs3 xus5 uxs6 declined, indicating that UDP-Xyl from cytosol AtUXS participates in xylan synthesis. Gel-permeation chromatography showed that the molecular weight of non-cellulosic polysaccharides in the triple mutants, mainly composed of xylans, is lower than that in the wild type, suggesting an effect on the elongation of the xylan backbone. Upon saccharification treatment stems of the uxs3 uxs5 uxs6 triple mutants released monosaccharides with a higher efficiency than those of the wild type. Taken together, our results indicate that the cytosol UXS plays a more important role than the Golgi-localized UXS in xylan biosynthesis.

Expedited extraction of xylan from corncob by power ultrasound

Some lignocellulosic biomass like corncob contains a large amount of xylan,a valuable material for applications in food,chemical and pharmaceutical industries.Conventional extraction of xylan is a time consuming process,requiring 12 to 24 hours to complete.In this study,the efficacy of power ultrasound for reducing the extraction time and increasing the xylan yield was investigated.Two acoustic energy densities(0.18 W/mL and 0.45 W/mL)of the power ultrasound were applied to corncob for 10,20,30,and 60 min.The treated samples were steamed at 110,118 and 121℃for 30 min.At 0.45 W/mL acoustic energy density and 121℃steaming temperature,ultrasonic treatment was able to extract 39%of xylan in 43 min.In contrast,the conventional treatment was able to extract only 34%of xylan in 24 h.Power ultrasound assisted extraction reduced the processing time by 97%in addition to yield increase by 14.7%.Power ultrasound can be a potential technology for enhanced xylan extraction from corncob to result in significant processing time savings.

Optimization of fermentation medium for xylanase-producing strain Xw2

To improve the fermentation yield of xylanase by optimizing the fermentation conditions for strain Xw2, a Plackett-Burman design was used to evaluate the effects of eight variables on xylanase production by strain Xw2. The steepest ascent （descent） method was used to approach the optimal response surface experimental area. The optimal fermentation conditions were obtained by central composite design and response surface analysis. The results showed that the composition of the optimal fermentation medium was corn cob ＋ 1.5% wheat bran （1：1）, 0.04% MnSO4, 0.04% K2HPO4. 3H2O, and an inoculum size of 6% in 50 mL liquid volume （pH = 6.0）. The optimal culture conditions were 28oc at 150 r/min for 54.23 h. The results of this study can serve as the basis for the industrial production and application of xylanase.