Screening and Statistical Optimization of Physiochemical Parameters for the Production of Xylanases from Agro-Industrial Wastes

Xylanases are mostly produced through submerged fermentation;nonetheless solid-state fermentation has increased profound attention and consideration of scholars having high conversion level biomass to energy conservation. This study depicted the purification of xylanases and their possible utilization in industry. The present study was carried out to examine the culture influence of fungal strain Fomes fomentarius (F. fomentarius) using different agro-industrial residues (wheat straw, rice husk, sugarcane bagasse and siris pods). F. fomentarius showed maximum enzyme production after 72 h of fermentation, when grown on wheat straw in solid state fermentation process while maximum activity showed on pH 6.0 at 30°C. The other parameters optimized by statistical design (RSM) showed maximum xylanase activity (146 ± 8 IU/mL) at 65% moisture content, 4 mL inoculums size, 175 mg Ammonium sulphate, 200 mg Calcium carbonate and 1.4 grams of glucose. Xylanase was salted out at 60% ammonium sulphate concentration and enzyme was further purified by Sephadex G-100 gel filtration chromatography with 2.2 fold increase in activity. The purified xylanase from F. fomentarius had optimum pH 6.0 and 40°C. Xylanase showed higher specificity for oat spelt xylan with kinetic constants Km 1.25 mg/mL and Vmax 54 mM/min. Xylanases have an industrial important enzyme used extensively in food, feed and paper industry.

Investigation of the nutritional and functional roles of a combinational use of xylanase and β-glucanase on intestinal health and growth of nursery pigs

Background Xylanase andβ-glucanase combination(XG)hydrolyzes soluble non-starch polysaccharides that are anti-nutritional compounds.This study aimed to evaluate the effects of increasing levels of XG on intestinal health and growth performance of nursery pigs.Methods Forty pigs(6.5±0.4 kg)were assigned to 5 dietary treatments and fed for 35 d in 3 phases(11,9,and 15 d,respectively).Basal diets mainly included corn,soybean meal,and corn distiller's dried grains with solubles,contained phytase(750 FTU/kg),and were supplemented with 5 levels of XG at(1)0,(2)280 TXU/kg xylanase and 125 TGU/kgβ-glucanase,(3)560 and 250,(4)840 and 375,or(5)1,120 and 500,respectively.Growth performance was measured.On d 35,all pigs were euthanized and jejunal mucosa,jejunal digesta,jejunal tissues,and ileal digesta were collected to determine the effects of increasing XG levels and XG intake on intestinal health.Results Increasing XG intake tended to quadratically decrease(P=0.059)viscosity of jejunal digesta(min:1.74 m Pa·s at 751/335(TXU/TGU)/kg).Increasing levels of XG quadratically decreased(P<0.05)Prevotellaceae(min:0.6%at 630/281(TXU/TGU)/kg)in the jejunal mucosa.Increasing XG intake quadratically increased(P<0.05)Lactobacillaceae(max:40.3%at 608/271(TXU/TGU)/kg)in the jejunal mucosa.Increasing XG intake quadratically decreased(P<0.05)Helicobacteraceae(min:1.6%at 560/250(TXU/TGU)/kg)in the jejunal mucosa.Increasing levels of XG tended to linearly decrease(P=0.073)jejunal Ig G and tended to quadratically increase(P=0.085)jejunal villus height to crypt depth ratio(max:2.62 at 560/250(TXU/TGU)/kg).Increasing XG intake tended to linearly increase the apparent ileal digestibility of dry matter(P=0.087)and ether extract(P=0.065).Increasing XG intake linearly increased(P<0.05)average daily gain.Conclusions A combinational use of xylanase andβ-glucanase would hydrolyze the non-starch polysaccharides fractions,positively modulating the jejunal mucosa-associated microbiota.Increased intake of these enzyme combination possibly reduced digesta viscosity and humoral immune response in the jejunum resulting in improved intestinal structure,and ileal digestibility of nutrients,and finally improving growth of nursery pigs.The beneficial effects were maximized at a combination of 550 to 800 TXU/kg xylanase and 250 to 360 TGU/kgβ-glucanase.

A Rapid Parameter of Enzyme-Treated Cellulosic Material Revealed by Reducing Sugar Release

This study was conducted to evaluate the effectiveness of enzymes in purifying and reducing the degree of polymerization of cellulose for the production of dissolving pulp.Our goal was to determine the contributions of xylanase(X)and endoglucanase(EG)in the treatment of pulp,specifically by quantifying the formation of soluble and insoluble reducing sugars using the dinitrosalycilic acid(DNS)test.Predominantly,the release of soluble reducing sugars(RSSol)was enhanced after xylanase treatment,while endoglucanase(EG)treatment led to changes in insoluble reducing sugars(RSIns).The maximum synergism was observed for RSIns when a high ratio of endoglucanase to xylanase(320EG:5X/g pulp)was used.The relative contribution of endoglucanase to RSins was determined to be 15.6%of the total reducing sugar.The viscosity of pulps treated with xylanase decreased only by 7%,whereas endoglucanase treatment significantly reduced viscosity by 45%.Modifications in the particle size were observed after pulp treatment with the combination of endoglucanase and xylanase.In summary,the DNS test is a rapid and effective method for evaluating the efficiency of enzyme treatments on pulps.The measurement of RSIns correlates with changes in pulp viscosity to different extents,providing valuable insights into the effectiveness of enzyme treatments.

Enzymology properties of two different xylanases and their impacts on growth performance and intestinal microflora of weaned piglets

The enzyme xylanase is more and more widely used in feed production, but different xylanase have different properties, mechanism and application effects. To provide a theoretical basis for choosing more suitable xylanase in feed production, we selected bacterial xylanase(BX), labeled enzyme A, and trichoderma xylanase(TX), labeled enzyme B, and studied the enzymology properties and application effects on growth performance and gut flora in weaned piglets. The results showed that the activity levels of both appear parabolic along with increasing pH or temperature, but the amplitude of enzyme activity changing curves and the pH/temperature of optimal activity level are different, where enzyme A has the optimal activity level at 50 ℃ with a pH value of 5.0. The optimal activity level of enzyme B was achieved at 70 ℃ with a pH around 6.0. Enzyme B suffered very little activity loss with moisture level at16% and temperature from 80℃ to 90 ℃. Enzyme A suffered a big drop in activity level when processed with high temperature from around 80 ℃ to 90 ℃, and it was even completely inactivated at 90 ℃,Enzyme A has very low activity level after being processed in acid environment, but enzyme B has minor changes in activity level with respect to changes in acid level, indicating significantly different enzymatic properties between the two different sources of xylanases. In feeding experiment, the control group, was fed the basal diet, and the BX group and TX group were fed basal diets supplemented with 0,01% bacterial and fungal xylanases, respectively. The results showed that ADG of the BX group and TX group increased by 3.25%(P> 0.05) and 8.22%(P < 0.05), respectively, and the feed conversion ratio decreased by 6.74% and 7.86%(P > 0.05), respectively compared with the control group; TX group had significantly higher(P < 0.05) ADG compared with BX group; BX group and TX group had significantly lower ileum Escherichia coli level than the control group, which were reduced by up to 12.98%(P < 0.05) and 11.68%(P < 0.05), respectively, but the ileal lactic acid bacteria levels were significantly increased by 16.21%(P < 0.01) and 27.02%(P < 0.01),respectively. There were no significant differences(P > 0.05) between BX group and TX group in terms of lactic acid bacteria E. coli level. We concluded that fungal xylanase(enzyme B) has better performances in improving weaned piglet growth and in increasing ileal lactic acid bacteria level compared with bacterial xylanase(enzyme A).

Impact of xylanases on gut microbiota of growing pigs fed corn-or wheat-based diets

This study investigated the effects of xylanase supplementations with cereal-based diets on nutrient digestibility and gut microbiota of growing pigs. A total of 96 individually penned pigs(initial BW = 22.7 ± 0.65 kg) were allotted to 12 treatments and subjected to a completely randomized block design experiment. Pigs in each treatment were fed an isocaloric wheat-based or corn-based diet with or without 1 of 5 types of xylanase supplements(XA, XB, XC, XD, XE). On d 42, all piglets were euthanized to obtain ileal and cecal digesta for microbial analysis, which involved high-throughput sequencing of the V1 - V3 regions of 16 S rRNA gene. Corn-and wheat-based diets differed(P < 0.05) in digestion characteristics. Dietary treatments affected the alpha-and beta-diversities of microbiota in the cecum but not in the ileum. The wheat-based diet increased(P < 0.05) alphadiversity and clustered separately(P < 0.05) compared with the corn-based diet. Wheat-based diet also promoted the relative abundance of genus(g.) Succinivibrio while corn-based diet promoted the proportion of family(f.) Veillonellaceae in the community. Among xylanases, only XC within the wheat-based diet altered(P < 0.05) the beta-diversity of the cecal microbiota compared with control. For each cereal-based diet and compared with the controls, xylanase treatments affected(P < 0.05) the proportions of 5 bacterial taxa in the ileum(f. Peptostreptococcaceae, order [o.]Streptophyta, f. Clostridiaceae, g. Clostridium and g. Streptococcus) and 8 in the cecum(g. Lactobacillus,g. Streptococcus, class [c.] Clostridia, f. Clostridiaceae, g. Megasphaera, g. Prevotella, g. Roseburia and f.Ruminococcaceae). Network analysis showed that across diets under control treatments, Bacteroidetes was the most influential phylum promoting cooperative relationships among members of the ileum and cecum microbiota. Xylanase treatment, however, reduced the influence of Bacteroidetes and promoted a large number of hub taxa majority of which belonged to the Firmicutes phylum. To maximize the efficiency of xylanase supplementation, our data suggest that xylanase C originated from Bacillus subtilis was more effective when applied to wheat-based diets, while xylanase A originated from Fusarium verticillioides was more beneficial when applied to corn-based diets.

Debranching enzymes decomposed corn arabinoxylan into xylooligosaccharides and achieved prebiotic regulation of gut microbiota in broiler chickens

Background Corn arabinoxylan(AX)is a complicated and multibranched antinutritional factor,thereby proving the use of endo-xylanase(EX)to be marginally valid.This study focused on specific types of AX-degrading enzymes(ADEs)to exert the synergy of debranching enzymes and track the prebiotic potential of enzymatic hydrolysates.This study investigated the effects of ADEs on the growth performance,intestinal histomorphology,absorption functions,changes in polysaccharide components,fermentation,and gut microbiota of broiler chickens.Five hundred seventysix five-day-old Arbor Acres male broiler chickens were randomly allocated into eight treatments with six replicates each.Corn basal diets supplemented with or without enzymes were fed for a 21-day period,specifically including EX,its compatible use with arabinofuranosidase(EXA)or ferulic acid esterase(EXF),and compound groups with the above three enzymes(XAF).Results Specific ADEs stimulated the jejunal villus height and goblet cell number and evidently decreased the crypt depth(P<0.05),while the ratio of ileal villus height to crypt depth was significantly increased in EXF(P<0.05).Maltase activities of ileal mucosa in XAF groups were extremely enhanced(P<0.01),and EX boosted the activity of Na+-K+ATPase in the small intestine(P<0.01).The insoluble AX concentrations comparatively lessened,thereby notably raising the sundry xylooligosaccharide(XOS)yield in the ileal chyme(P<0.05),which was dominant in xylobiose and xylotriose.Improvements in the abundance and diversity of ileal microbial communities within the EXA,EXF,and XAF treatments were observed(P<0.05).Positive correlations between microbiota and XOS were revealed,with xylobiose and xylotriose being critical for ten beneficial bacteria(P<0.05).EXF increased the BWG and FCR of broiler chickens in this phase(P<0.05),which was attributed to the thriving networks modified by Lactobacillus.The intracecal contents of acetic acid,butyric acid,and propionic acid were greatly enhanced in most ADE groups,such as EXF(P<0.05).Conclusions Debranching enzymes appreciably targeted corn AX to release prebiotic XOS in the posterior ileum and facilitated intracaecal fermentation.It was beneficial for improving gut development,digestion and absorption and modulating the microflora to promote the early performance of broiler chickens.

Xylanase improves the intestinal barrier function of Nile tilapia(Oreochromis niloticus)fed with soybean(Glycine max)meal

Background Soybean(Glycine max)meal is one of the important protein sources for fish,but the non-starch polysaccharides(NSP)in soybean meal impair the intestinal barrier function.Here we aimed to investigate whether xylanase can alleviate the adverse effects on the gut barrier induced by soybean meal in Nile tilapia and to explore the possible mechanism.Results Nile tilapia(Oreochromis niloticus)(4.09±0.02 g)were fed with two diets including SM(soybean meal)and SMC(soybean meal+3,000 U/kg xylanase)for 8 weeks.We characterized the effects of xylanase on the gut barrier,and the transcriptome analysis was performed to investigate the underlying mechanism.Dietary xylanase improved intestinal morphology and decreased the concentration of lipopolysaccharide(LPS)in serum.The results of transcriptome and Western blotting showed that dietary xylanase up-regulated the expression level of mucin2(MUC2)which may be related to the inhibition of protein kinase RNA-like endoplasmic reticulum kinase(perk)/activating transcription factor 4(atf4)signaling pathways.Microbiome analysis showed that addition of xylanase in soybean meal altered the intestinal microbiota composition and increased the concentration of butyric acid in the gut.Notably,dietary sodium butyrate was supplemented into the soybean meal diet to feed Nile tilapia,and the data verified that sodium butyrate mirrored the beneficial effects of xylanase.Conclusions Collectively,supplementation of xylanase in soybean meal altered the intestinal microbiota composition and increased the content of butyric acid which can repress the perk/atf4 signaling pathway and increase the expression of muc2 to enhance the gut barrier function of Nile tilapia.The present study reveals the mechanism by which xylanase improves the intestinal barrier,and it also provides a theoretical basis for the application of xylanase in aquaculture.

Effect of Wheat Based Diet Supplemented with Xylanase on Blood Sugar and Total Protein in Serum of Geese

[Objective] The effects of wheat based diet supplemented with xylanase on blood sugar and total protein in serum of geese were studied. [ Method ] By using the randomized design of single factor, the 1-day-old healthy goslings were divided into 6 groups and fed with corn based diet, wheat based diet and wheat based diet supplemented with xylanase at different concentrations respectively, the contents of blood sugar and total protein in serum were determined. [ Result] The wheat based diet supplemented with xylanase could increase the blood sugar and total protein in serum of geese and wheat based diet supplemented with 0.2% xylanase generated the best effect, which was higher than those of corn based diet group. As for the concentration of protein in serum, wheat based diet supplemented with 0.2% xylanase was significantly different from corn based diet and wheat based diet. [ Conclusion] The wheat based diet supplemented with xylanase could enhance geese production.

利用真菌纤维素结合域(CBD)保守性序列进行草菇木聚糖酶cDNA的克隆

Cellulose binding domains (CBDs) are present in the majority of fungal cellulases and hemicellulases. Based on the conserved region of CBDs, degenerate primers were designed and used to amplify the 5′ end cDNA fragment of xylanase from Volvariella volvacea by 5′ RACE. Gene specific primer was then designed based on extreme region of 5′ end cDNA fragment and used to amplify the full length cDNA of xylanase. The cDNA of xyn 1 was 1 287 bp in length, including 3′ and 5′ non coding region. The xyn 1 cDNA contained an ORF of 1101 bp encoding 367 amino acids, in which there was a putative signal peptide with 19 amino acids. Alignment of the deduced amino acid sequence of xyn 1 with other xylanases showed that the homology with family 10 xylanases from Agaricus bisporus xyl1,Aspergillus sojae xyn1, Aspergillus kawachii xynA, Fusarium oxysporum f.sp xyl3 was 64%,55%,52%,55%, respectively.

Effect of Ultrasound and Xylanase Treatment on the Physical-Mechanical Properties of Bleached Eucalyptus Kraft Pulp

The modification on the fiber structure of bleached eucalyptus kraft pulp is a very attractive alternative for improve- ments in the properties of paper production. The enzymatic treatment by xylanases and ultrassonic treatments modify the characteristics of the fibers, has been reported. Therefore, the purpose of this study was to evaluate the influence of ultrasonic waves as a facilitator of the action of enzymes (hemicellulase) by modifying the physicochemical nature of fiber eucalyptus Kraft pulp, in order to improve the physical and mechanical properties of the paper. But it was observed the that idea of ultrasound acts as facilitator for action of enzymes can’t be affirmed, since in most properties XA-1 and XA-2 were equal statistically. It may be noted that the junction of ultrasound and xylanase provided improves on tensile index, specific elastic modulus and tensile energy absorption and a decrease of tear index on the mechanical properties of handsheet and it increased the opacity when the ultrasound was applied before xylanase.

Screening of a Composite Microbial System and Its Characteristics of Wheat Straw Degradation

To accelerate the decomposition of wheat straw directly returned to soil, we constructed a microbial system （ADS-3） from agricultural soil containing rotting straw residues using a 40-wk limited cultivation. To assess its potential use for accelerating straw decomposing, the decomposing characteristics and the microbial composition of ADS-3 were analyzed. The results indicated that it could degrade wheat straw and filter paper by 63.8 and 80%, respectively, during 15 d of incubation. Straw hemicellulose degraded dramatically 51.2% during the first 3 d, decreasing up to 73.7% by the end of incubation. Cellulose showed sustained degradation reaching 53.3% in 15 d. Peak values of xylanase and cellulase activities appeared at 3 and 11 d, with 1.32 and 0.15 U mL-1, respectively. Estimated pH averaged 6.4-7.6 during the degradation process, which approximated acidity and alkalinity of normal soils. The microbial composition of ADS-3 was stable based on denaturing gradient gel electrophoresis （DGGE） analysis. By using bacterial 16S rRNA and fungal 26S rRNA gene clone library analysis, 20 bacterial clones and 7 fungal clones were detected. Closest identified relatives of bacteria represented by Bacillus fusiformis, Cytophaga sp., uncultured Clostridiales bacterium, Ruminobacillus xylanolyticum, Clostridium hydroxybenzoicum, and uncultured proteobacterium and the fungi were mainly identified as related to Pichia sp. and uncultured fungus.

Studies on Mutation Breeding of High-Yielding Xylanase Strains by Low-Energy Ion Beam Implantation

As a new mutagenetic method, low-energy ion implantation has been used widely in many research areas in recent years. In order to obtain some industrial strains with high xylanase yield, the wild type strain Aspergillus niger A3 was mutated by means of nitrogen ions implantation （10 keV, 2.6× 10^14 ～ 1.56 × 10^15 ions/cm^2） and a mutant N212 was isolated subsequently. However, it was found that the initial screening means of the high-yielding xylanase strains such as transparent halos was unfit for first screening. Compared with that of the wild type strain, xylanase production of the mutant N212 was increased from 320 IU/ml to 610 IU/ml, and the optimum fermentation temperature was increased from 28 ℃ to 30 ℃.

Effects of cellulase and xylanase enzymes mixed with increasing doses of Salix babylonica extract on in vitro rumen gas production kinetics of a mixture of corn silage with concentrate

An in vitro gas production （GP） technique was used to investigate the effects of combining different doses of Salix babylonica extract （SB） with exogenous fibrolytic enzymes （EZ） based on xylanase （X） and cellulase （C）, or their mixture （XC; 1：1 v/v） on in vitro fermentation characteristics of a total mixed ration of corn silage and concentrate mixture （50：50, w/w） as substrate. Four levels of SB （0, 0.6, 1.2 and 1.8 mL g-1 dry matter （DM）） and four supplemental styles of EZ （1 μL g-1 DM; control （no enzymes）, X, C and XC （1：1, v/v） were used in a 4×4 factorial arrangement. In vitro GP （mL g-1 DM） were recorded at 2, 4, 6, 8, 10, 12, 24, 36, 48 and 72 h of incubation. After 72 h, the incubation process was stopped and supernatant pH was determined, and then filtered to determine dry matter degradability （DMD）. Fermentation parameters, such as the 24 h gas yield （GY24）, in vitro organic matter digestibility （OMD）, metabolizable energy （ME）, short chain fatty acid concentrations （SCFA）, and microbial crude protein production （MCP） were also estimated. Results indicated that there was a SBxEZ interaction （P〈0.0001） for the asymptotic gas production （b）, the rate of gas production （c）, GP from 6 to 72 h, GP2 （P=0.0095）, and GP4 （P=0.02）. The SB and different combination of enzymes supplementation influenced （P〈0.001） in vitro GP parameters after 12 h of incubation; the highest doses of SB （i.e., 1.8 mL g-1 DM）, in the absence of any EZ, quadratically increased （P〈0.05） the initial delay before GP begins （L） and GP at different incubation times, with lowering b （quadratic effect, P〈0.0001 ） and c （quadratic effect, P〈0.0001 ; linear effect, P=0.0018）. The GP was the lowest （P〈0.05） when the highest SB level was combined with cellulose. There were SBxEZ interactions （P〈0.001） for OMD, ME, the partitioning factor at 72 h of incubation （PF72）, GY24, SCFA, MCP （P=0.0143）, and pH （P=0.0008）. The OMD, ME, GY24 and SCFA with supplementation of SB extract at 1.8 mL g-1 DM were higher （P〈0.001） than the other treatments, however,PF72 was lower （quadratic effect, P=0.0194） than the other levels. Both C and X had no effect （P〉0.05） on OMD, pH, ME, GY24, SCFA and MP. The combination of SB with EZ increased （P〈0.001） OMD, ME, SCFA, PFz2 and GP24, whereas there was no impact on pH. It could be concluded that addition of SB extract, C, and X effectively improved the in vitro rumen fermentation, and the combination of enzyme with SB extract at the level of 1.2 mL g-1 was more effective than the other treatments.

Mutation-Screening in Xylanase-Producing Strains by Ion Implantation

With ion implantation (N+, energy 10 keV and dosage 1.56×1015 N+cm-2), a high xylanase-producing strain Aspergillus niger N212 was selected. Based on an orthogonal experiment, an optimal fermentation condition was designed for this high-yield strain. The suitable medium was composed of 8% corncob; 1.0% wheat bran; 0.1%TWEEN20; 0.5% (NH4)2SO4; 0.5%NaNO3; 0.5%FeSO4, 7.5 × 10-4; MnSO4·H2O, 2.5 × 10-4; ZnSO4, 2.0 × 10-4; CoCl2, 3.0 × 10-4. At present, under our experiment condition, xylanase activity of Aspergillus niger N212 reached a level of 600 IU/ml, almost increased by 100% in xylanase production and the time of yielding xylanase was largely reduced to 12 h at 28℃.

Thermostable ethanol tolerant xylanase from a cold-adapted marine species Acinetobacter johnsonii

A xylanase-producing bacterium, isolated from deep sea sediments, was identified as the cold-adapted marine species Acinetobacter Johnsonii. A cold-adapted marine species Acinetobacter Johnsonii could grow at 4 ℃. The optimum temperature and pH of xylanase from a cold-adapted marine species Acinetobacter Johnsonii were 55 ℃ and pH 6.0. Xylanase from a cold-adapted marine species Acinetobacter Johnsonii remained at 80% activity after incubation for 1 h at 65 ℃. The xylanase activity was 1.2-fold higher in 4% ethanol solution than in ethanol free solution. Gibbs free energy of denaturation, ΔG, was higher in 4% ethanol solution than in ethanol free solution. Thermostable ethanol tolerant xylanase was valuable for bioethanol production by simultaneous saccharification and fermentation process with xylan as a carbon source.

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.

Improvement of xylanase production by Aspergillus niger XY-1 using response surface methodology for optimizing the medium composition

Objective: To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation (SSF). Methods: Statistical methodology including the Plackett-Burman design (PBD) and the central composite design (CCD) was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results: Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and (NH4)2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization. Finally, the CCD and response surface methodology (RSM) were applied to determine the optimal concentration of each sig-nificant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by multiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows: x1 (urea)=0.163 (41.63 g/L), x2 (Na2CO3)=?1.68 (2.64 g/L), x3 (MgSO4)=1.338 (10.68 g/L) and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion: By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization.

Mutagenesis and Screening of High Yield Xylanase Production Strain of Aspergillus usamii by Microwave Irradiation

A high yield xylanase producing strain, A. usamii L336-23, was screened out from its parent strain A.usamii L336 after microwave irradiation. Its productivity of xylanase activity increased by 35.7% from 21000μu·ml-1 to 28500μu·ml-1 and was stable after frequent subcultures and storage for more than two months. The mechanism of microwave mutation was also discussed.

Expression and Characterization of a Thermostable Xylanase Gene xynA from a Themophilic Fungus in Pichia pastoris

The gene of xylanase （xynA） was amplified by RT-PCR from the total RNA of a themophilic fungus Thermomyces lanuginosus SY2. The sequence analysis showed that gene coding region of mature peptide contained 0.585 kb, which coded 194 amino acids. The putative amino acid sequence and DNA sequence of xylanase from T. lanuginosus SY2 （GenBank no.： GU166389） were 98.97 and 99.49% identical to the other T. lanuginosus （GenBank no.： U35436）. A recombinant plasmid pPIC9K-xynA was constructed by inserting gene xynA into Pichia pastoris secretory vector pPIC9K. Linearized pPIC9K-xynA was transformed into P. pastoris GS115 with the method of electroporation. The recombinant strain was identified by G418 selection and confirmed by PCR analysis. It was induced by 1.0% methanol at 28°C to express the recombinant xylanase. The results showed that the recombinant xylanase was secreted into extracellular fermentation liquid. The highest enzyme activity of 113.5 IU mL-1 and protein content of 889.7 μg mL-1 were detected for 216 h of induction. The optimal pH value and temperature of the enzyme activity was 5.5 and 65°C, respectively. The xylanase activity retained above 80% from pH value 2.5 to 8.5 for 48 h. The enzyme activity was above 85% at incubation temperature of 55°C.

Optimization of Fermentation Conditions for Xylanase Production by Aspergillus niger NS-1

In this study, a xylanase-produeing Aspergillus niger strain, NS-1, was screened and isolated from agricultural and forestry wastes. Based on single-fac- tor experiments, the effects of different carbon sources, composite carbon sources, nitrogen sources, calcium carbonate concentrations, initial pH and surfactants on xylanase production by A. niger NS-1 were investigated. The results indicated that the most appropriate carbon source was corncobs ; the best composite carbon source was corncobs ＋ xylan, which was conducive to xylanase secretion; the most suitable nitrogen source was ammonium sulfate. Xylanase activity reached the highest in the medium added with 1.5% calcium carbonate and SDS as a surfactant with an initial pH of 5.0. This study provided the basis for the production of high-activity xylanase.