• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.638-644
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• 2018

In this study, the immobilization of xylanase using a protein-inorganic hybrid nanoflower system was assessed to improve the enzyme properties. The synthesis of hybrid xylanase nanoflowers was very effective at $4^{\circ}C$ for 72 h, using 0.25 mg/ml protein, and efficient immobilization of xylanase was observed, with a maximum encapsulation yield and relative activity of 78.5% and 148%, respectively. Immobilized xylanase showed high residual activity at broad pH and temperature ranges. Using birchwood xylan as a substrate, the $V_{max}$ and $K_m$ values of xylanase nanoflowers were 1.60 mg/ml and $455{\mu}mol/min/mg$ protein, compared with 1.42 mg/ml and $300{\mu}mol/min/mg$ protein, respectively, for the free enzyme. After 5 and 10 cycles of reuse, the xylanase nanoflowers retained 87.5% and 75.8% residual activity, respectively. These results demonstrate that xylanase immobilization using a proteininorganic hybrid nanoflower system is an effective approach for its potential biotechnological applications.

• Microbiology and Biotechnology Letters
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• v.43 no.4
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• pp.330-335
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• 2015

The optimum conditions for the production of xylanase from Bacillus agaradhaerens DK-2386 have been previously investigated. In this study xylanase was purified by ammonium sulfate precipitation and CM-sepharose ion exchange chromatography. The molecular mass of the xylanase as determined by SDS-PAGE was 23 kDa in a form of monomeric enzyme. The optimum pH and temperature for xylanase activity was 6.0 and $60^{\circ}C$, respectively. Xylanase activity was increased by the addition of EDTA and then stabilized at $40^{\circ}C$ for 24 h. The maximum xylanase activity was obtained when Birchwood xylan was used as a substrate and the $V_{max}$ and $K_m$ were $49,724{\mu}mol/min$ and 6.08 mg/ml, respectively.

• Journal of the Korean Wood Science and Technology
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• v.46 no.6
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• pp.670-680
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• 2018

Acanthophysium sp. KMF001, a wood rotting fungus, produces a strong crude enzyme complex that efficiently produces simple sugars from wood. The transcriptomic analysis of Acanthophysium sp. KMF001 identified 14 genes for putative glycoside hydrolases. Among them, isotig01043 was expressed heterogeneously in Escherichia coli BL21(DE3), and the expressed protein exhibited an endo-${\beta}$-1,4-xylanase activity which showed the optimum reaction at pH 5.0 and $30^{\circ}C$. The enzyme kinetic values of $K_m$ and $V_{max}$ were 25.92 mg/ml and $0.628{\mu}mole/mg/ml$, respectively. The enzymatic characteristics of the expressed xylanase showed a typical fungal xylanase. However, the bioinformatics analysis suggested that the protein encoded by isotig01043 was a novel xylanase based on a low identity when it was compared with the closest protein in the NCBI database and a similar protein domain with GH16_fungal_Lam16A_glucanase, which had not been earlier suggested as a xylanase.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.1
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• pp.57-63
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• 2017

Objective: Two experiments were conducted to determine the effects of adding exogenous phytase and xylanase, individually or in combination, as well as pelleting on nutrient digestibility, available energy content of wheat and the performance of growing pigs fed wheat-based diets. Methods: In Experiment 1, forty-eight barrows with an initial body weight of $35.9{\pm}0.6kg$ were randomly assigned to a $2{\times}4$ factorial experiment with the main effects being feed form (pellet vs meal) and enzyme supplementation (none, 10,000 U/kg phytase, 4,000 U/kg xylanase or 10,000 U/kg phytase plus 4,000 U/kg xylanase). The basal diet contained 97.8% wheat. Pigs were placed in metabolic cages for a 7-d adaptation period followed by a 5-d total collection of feces and urine. Nutrient digestibility and available energy content were determined. Experiment 2 was conducted to evaluate the effects of pelleting and enzymes on performance of wheat for growing pigs. In this experiment, 180 growing pigs ($35.2{\pm}9.0kg\;BW$) were allocated to 1 of 6 treatments according to a $2{\times}3$ factorial treatment arrangement with the main effects being feed form (meal vs pellet) and enzyme supplementation (0, 2,500 or 5,000 U/kg xylanase). Results: In Experiment 1, there were no interactions between feed form and enzyme supplementation. Pelleting reduced the digestibility of acid detergent fiber (ADF) by 6.4 percentage units (p<0.01), increased the digestibility of energy by 0.6 percentage units (p<0.05), and tended to improve the digestibility of crude protein by 0.5 percentage units (p = 0.07) compared with diets in mash form. The addition of phytase improved the digestibility of phosphorus (p<0.01) and calcium (p<0.01) by 6.9 and 7.6 percentage units respectively compared with control group. Adding xylanase tended to increase the digestibility of crude protein by 1.0 percentage units (p = 0.09) and increased the digestibility of neutral detergent fiber (NDF) (p<0.01) compared with control group. Supplementation of the xylanase-phytase combination improved the digestibility of phosphorus (p<0.01) but impaired NDF digestibility (p<0.05) compared with adding xylanase alone. In Experiment 2, adding xylanase increased average daily gain (p<0.01) and linearly improved the feed:gain ratio (p<0.01) compared with control group. Conclusion: Pelleting improved energy digestibility but decreased ADF digestibility. Adding xylanase increased crude protein digestibility and pig performance. Phytase increased the apparent total tract digestibility of phosphorus and calcium. The combination of phytase-xylanase supplementation impaired the effects of xylanase on NDF digestibility.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.153-156
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• 2002

The characterization of a partially purified, cellulase-free, thermostable alkaline xylanase from thermoalkalophilic Bacillus sp. JB-99 was investigated. The xylanase production was the highest when birchwood xylan was added to a medium containing finely powdered rice bran, showing 4,826 IU$ml^-1$ of activity for 15 h of incubation. The partially purified xylanase exhibited an optimum temperature and pH at $70^C{\circ}$ and 10, respectively. The enzyme was stable at pH 5-11 at $50^C{\circ}$. The xylanase activity was strongly inhibited by $Hg^2+$, while dithiothreitol, cysteine, and ${\beta}$-mercaptoethanol enhanced the activity.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.9
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• pp.1491-1499
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• 2018

Objective: This study was designed to investigate the effects of an Aspergillus sulphureus xylanase expressed in Pichia pastoris on the growth performance, nutrient digestibility and gut microbes in weanling pigs. Methods: A total of 180 weanling pigs (initial body weights were $8.47{\pm}1.40kg$) were assigned randomly to 5 dietary treatments. Each treatment had 6 replicates with 6 pigs per replicate. The experimental diets were wheat based with supplementation of 0, 500, 1,000, 2,000, and 4,000 U xylanase/kg. The experiment lasted 28 days (early phase, d 0 to 14; late phase, d 15 to 28). Results: In the early phase, compared to the control, average daily gain (ADG) was higher for pigs fed diets supplemented with xylanase and there was a quadratic response in ADG (p<0.05). In the entire phase, ADG was higher for the pigs fed 1,000 or 2,000 U/kg xylanase compared to the control (p<0.05). The gain to feed ratio was higher for pigs fed diets supplemented with 1,000 or 2,000 U/kg xylanase compared to the control (p<0.05). Increasing the amount of xylanase improved the apparent total tract digestibility of dry matter, crude protein, neutral detergent fiber, calcium, and phosphorus during both periods (p<0.05). Xylanase supplementation (2,000 U/kg) decreased the proportion of Lachnospiraceae (by 50%) in Firmicutes, but increased Prevotellaceae (by 175%) in Bacteroidetes and almost diminished Enterobacteriaceae (Escherichia-Shigella) in Proteobacteria. Conclusion: Xylanase supplementation increased growth performance and nutrient digestibility up to 2,000 U/kg. Supplementation of xylanase (2,000 U/kg) decreased the richness of gut bacteria but diminished the growth of harmful pathogenic bacteria, such as Escherichia-Shigella, in the colon.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.11
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• pp.1659-1664
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• 2008

To study the working mechanisms for non-starch polysaccharidases to improve the growth performance of broiler chickens, a 21-day feeding trial was conducted. Two dietary treatments were included: 1) wheat diet (the control); 2) wheat+xylanase diet (xylanase, Allzyme PT, Alltech, Kentucky, USA). There were 8 replicates with 8 birds each for each treatment and the experimental diets were given to birds from hatch. Feed intake and body weight were measured on days 7 and 21. At the same ages, samples were taken for the determination of selected groups of luminal and mucosa-associated bacteria, mucosal morphology, brush-border membrane (BBM) bound enzyme activity and ileal nutrient digestibility. The xylanase supplement increased (p<0.05) body weight gain (BWG) and improved feed conversion ratio (FCR) at the end of the experiment but protein and starch digestibilities were not affected (p>0.05) by xylanase. Up to day 7, xylanase increased the counts of C. perfringens in the ileum and total anaerobic bacteria (TAB) in the caeca (p<0.05, p=0.07, respectively). By day 21, the counts of ileal lactobacilli (p<0.05) and TAB (p=0.07) were lower in birds given the xylanase-supplemented diet than in those on the control diet. No significant differences were observed in the counts of mucosa-associated lactobacilli and coliforms between xylanase treatment and the control at both ages. Villus height at the jejunum was not affected (p>0.05) by the supplement but crypt depth at the same site was reduced at day 7. Also, xylanase tended to increase the concentration of BBM protein (p = 0.09) and the specific activity of sucrase (p = 0.07) at day 21.

• Journal of Animal Science and Technology
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• v.63 no.6
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• pp.1275-1285
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• 2021

The objective of the present experiment was to investigate the effect of dietary palm kernel meal (PKM) and β-xylanase supplementation on productive performance, egg quality, fatty liver incidence, and excreta characteristics in laying hens. A total of 320 Hy-Line Brown laying hens (33 weeks of age) were allotted to 1 of 4 treatments with 8 replicates in a feeding trial. Each replicate consisted of 10 consecutive cages with 1 hen per cage. The corn-soybean meal-based control diet was prepared. Additional diet was prepared by including 10% of PKM in the control diet with a partial replacement of corn, soybean meal, and animal fat. In addition, 0.025% β-xylanase was supplemented at the expense of celite to those 2 diets to produce 4 treatment diets in a 2 × 2 factorial arrangement. All hens were provided the diet and water ad libitum for 8 weeks. Results indicated no significant interactions between inclusion of dietary PKM and β-xylanase for all measurements; therefore, the main effects were mainly discussed. Hens fed diets containing 10% PKM had greater (p < 0.05) feed intake and yolk color than those fed diets containing no PKM. However, dietary PKM did not influence fatty liver incidence and excreta characteristics. Dietary β-xylanase supplementation had no effects on all measurements, regardless of inclusion of PKM. In conclusion, PKM can be a potential feed ingredient for laying hens at the inclusion of 10% in the diet. It appears that dietary β-xylanase used in the current experiment has little effect on layer productivity, regardless of inclusion of 10% PKM in the diet.

• Journal of Life Science
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• v.22 no.7
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• pp.912-919
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• 2012

A Bacillus sp. strain producing celluase and xylanase was isolated from environmental soil with LB agar plate containing carboxymethylcellulose (CM-cellulose) and beechwood xylan stained with trypan blue as substrates, respectively. Based on the 16S rRNA gene sequence and API 50 CHL test, the strain was identified as B. subtilis and named B. subtilis NC1. The cellulase and xylanase from B. subtilis NC1 exhibited the highest activities for CM-cellulose and beechwood xylan as substrate, respectively, and both enzymes showed the maximum activity at pH 5.0 and $50^{\circ}C$. We cloned and sequenced the genes for cellulase and xylanase from genomic DNA of the B. subtilis NC1 by the shot-gun cloning method. The cloned cellulase and xylanase genes consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with a calculated molecular mass of 55,251 Da and a 1,269 bp ORF encoding a 422 amino acid protein with a calculated molecular mass of 47,423 Da, respectively. The deduced amino acid sequences from the genes of cellulase and xylanase showed high identity with glycosyl hydrolases family (GH) 5 and 30, respectively.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.527-533
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• 1992

The optimal conditions for cellulase and xylanase production by Trichoderma reesei 28217 were studied for enzymatic deinking of old newspaper. The amounts of cellulase and xylanase from the strain was varied by initial medium pH, Tween 80, inoculum size of spore suspension, and carbon and nitrogen sources. The optimal conditions for cellulase production were pH 5.0-6.5, 0.02% of Tween 80, 0.5-1.0% of inoculum size of spore suspension ($1{\times}10^{7}$/ml). cottonseed meal as nitrogen source, and corn flour as carbon source. On the other hand, the optimal conditions for xylanase production were pH 6.5, 0.01% of Tween 80, corn steep liquor as nitrogen source, and disintegrated old newspaper as carbon source. The inoculum size for xylanase production was the same as for cellulase production. The concomitant production of cellulase and xylanase in shake flask culture was efficiently induced in the medium containing 0.5% cottonseed meal as nitrogen source and 1.0% old newspaper and 2.0% corn flour as carbon sources. In this case the activities of cellulase and xylanase produced were 6.11-7.22 IU/mJ and 97.7 IU/ml. respectively. However, the cellulase production in $5{\ell}$ fermentor scale was slightly decreased compared with that in flask scale. Moreover, xylanase production was severely reduced in a fermentor scale. The study for the reason of decreased enzyme production in fermentor is further needed.