• Microbiology and Biotechnology Letters
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• v.41 no.3
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• pp.263-271
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• 2013

A xylanase-producing bacterium was isolated from a soil sample collected in Yongin city, Korea. The strain was aerobic and gram positive, and grew between pH 5.0 and 11.0, forming a yellow-colored colony. The strain was classified as a novel subspecies bacterium of Paenibacillus barcinonensis by 16S rRNA gene sequence similarity, phylogenetic analysis, phenotypic, and biochemical characteristics, and thus named Paenibacillus sp. HX-1. This strain produced extracellular endo-${\beta}$-1,4-xylanase, and the best xylanolytic activity (205.17 unit/ml) was obtained at 96 h in an optimized TNX medium containing 1% (w/v) bacto tryptone, 1% (w/v) NaCl, and 0.7% (w/v) beechwood xylan at pH 7.0, $37^{\circ}C$ and 200 rpm. The endo-${\beta}$-1,4-xylanase produced by the strain HX-1 yielded xylobiose as the end product from beechwood xylan hydrolysis. The enzyme exhibited optimum pH and temperature at pH 7.0 and $45^{\circ}C$, respectively. The remarkable enhancing effect of the TNX medium on xylanase production by HX-1, in spite of its simple formula, may have great advantages for industrial applications of xylanase.

• Current Research on Agriculture and Life Sciences
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• v.16
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• pp.45-54
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• 1998

A thermotolerant bacterium, Streptomyces thermocyaneoviolaceus which produced xylan-degrading enzymes, utilized excellently xylan of wheat bran by producing the enzymes in comparison with that of birchwood or oat spelts. Optimal enzyme production was achieved in WB medium containing 0.8% wheat bran, 0.06% yeast extract, 0.06% bactopeptone, 0.05% $MgSO_4{\cdot}7H_2O$, 0.05% $FeSO_4{\cdot}7H_2O$, 0.05% $KH_2PO_4$ and, 0.2% $K_2HPO_4$(pH 7.0) at $50^{\circ}C$ for 24 hrs. The optimal pH and temperature for the hydrolysis of xylan were pH 5.5 and $65^{\circ}C$, respectively. The enzyme activity was retained more than 80% at the range from pH 4.5 to pH 9.5 at $4^{\circ}C$ for 12 hrs and 94% on the heat-treatment at $65^{\circ}C$ for 1 hr. Xylobiose, xylotriose, xylose, and other xylooligosaccharides were produced as end products from hydrolysis of birchwood xylan by the xylanase of S. thermocyaneoviolceus.

• Proceedings of the Microbiological Society of Korea Conference
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• 2003.05a
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• pp.122-124
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• 2003

Cyclodextrin glucanotransferase (CGTase) and endoxylanase genes of Bacillus sp. were subcloned down-stream of yeast ADH1 promoter and expressed in S. cerevisiae. Most of the CGTase and endoxylanase expressed were detected in the extracellular medium with activity of 0.6 and 7-8 unit/ml, respectively. The recombinant enzymes were secreted as N-linked-glycosylated forms, resulting an enhanced thermal stability. CGTase predominantly produced $\alpha-cyclodextrin$ from starch and endoxylanase produced xylobiose and xylotriose from xylan.

• Journal of Microbiology and Biotechnology
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• v.27 no.4
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• pp.775-784
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• 2017

A neutral xylanase (CcXyn) was identified from Coprinus cinereus. It has a single GH10 catalytic domain with a basic amino acid-rich extension (PVRRK) at the C-terminus. In this study, the wild-type (CcXyn) and C-terminus-truncated xylanase ($CcXyn-{\Delta}5C$) were heterologously expressed in Pichia pastoris and their characteristics were comparatively analyzed with aims to examine the effect of this extension on the enzyme function. The circular dichorism analysis indicated that both enzymes in general had a similar structure, but $CcXyn-{\Delta}5C$ contained less ${\alpha}-helices$ (42.9%) and more random coil contents (35.5%) than CcXyn (47.0% and 32.8%, respectively). Both enzymes had the same pH (7.0) and temperature ($45^{\circ}C$) optima, and similar substrate specificity on different xylans. They all hydrolyzed beechwood xylan primarily to xylobiose and xylotriose. The amounts of xylobiose and xylotriose accounted for 91.5% and 92.2% (w/w) of total xylooligosaccharides (XOS) generated from beechwood by CcXyn and $CcXyn-{\Delta}5C$, respectively. However, truncation of the C-terminal 5-amino-acids extension significantly improved the thermostability, SDS resistance, and pH stability at pH 6.0-9.0. Furthermore, $CcXyn-{\Delta}5C$ exhibited a much lower $K_m$ value than CcXyn (0.27 mg/ml vs 0.83 mg/ml), and therefore, the catalytic efficiency of $CcXyn-{\Delta}5C$ was 2.4-times higher than that of CcXyn. These properties make $CcXyn-{\Delta}5C$ a good model for the structure-function study of $({\alpha}/{\beta})_8$-barrel-folded enzymes and a promising candidate for various applications, especially in the detergent industry and XOS production.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.147-152
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• 2009

A xylanase was purified from the culture supernatant of Bacillus sp. A-6 by using ultrafiltration and ion exchange chromatography on the column of SP-Sepharose using 5 mM acetate buffer, pH 5.0. The xylanase was eluted from the column at the concentration less than 0.05 M NaCl. The eluted xylanase was shown to be a single protein band in SDS-PAGE. Zymogram analysis indicated that the protein band in SDS-PAGE had the enzyme activity to hydrolyze oat spelt xylan. The molecular weights of the xylanase were 15,000 based on SDS-PAGE and 14,100 based on gel filtration chromatography. Thin layer chromatography showed that the xylanase hydrolyzed oat spelt xylan into xylobiose and high-molecular-weight xylooligosaccharides. The relative activities of the heated xylanase decreased to 80% at $40^{\circ}C$ after 7 hr and less than 40% at $60^{\circ}C$ after 1 hr.

• Journal of Life Science
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• v.20 no.12
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• pp.1801-1806
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• 2010

A $\beta$-xylosidase encoding gene from Klebsiella sp. Sc was cloned in Escherichia coli. The $\beta$-xylosidase gene consisted of an open reading frame of 1,680 nucleotides and encodes 559 amino acids with a deduced molecular weight of 63 kDa. The deduced amino acid sequence of the $\beta$-xylosidase from Klebsiella sp. Sc exhibits 90% identities and 95% positives compared to those from Klebsiella oxytoca (KOX), Lactobacillus lactis (LAC, 82%, 90%), Bacillus longum (BLON, 69%, 81%) and Escherichia coli (ECOLI, 47%, 63%). The $\beta$-xylosidase was purified by GST-fusion purification system. The pH and temperature optima of the enzyme were 6.6 and $55^{\circ}C$, respectively. The $\beta$-xylosidase hydrolyzes xylobiose to xylose.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1476-1484
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• 2015

Three endoxylanase-encoding genes from the moderately themophilic chemoorganotrophic bacterium Melioribacter roseus were cloned and expressed in Escherichia coli. Genes xyl2091 (Mros_2091) and xyl2495 (Mros_2495) encode GH10 family hydrolases, whereas xyl2090 (Mros_2090) represents the GH30 family. In addition to catalytic domains, Xyl2090 and Xyl2091 contain carbohydrate-binding modules that could facilitate their binding to xylans and Por sorting domains associated with the sorting of proteins from the periplasm to the outer membrane, where they are covalently attached. Recombinant endoxylanase Xyl2495 exhibited a high specific activity of 1,920 U/mg on birchwood xylan at 40℃. It is active at low temperatures, exhibiting more than 30% of the maximal activity even at 0℃. Endoxylanases Xyl2090 and Xyl2091 have lower specific activities but higher temperature optima at 80℃ and 65℃, respectively. Analysis of xylan hydrolysis products revealed that Xyl2090 generates xylo-oligosaccharides longer than xylopentaose. Xylose and xylobiose are the major products of xylan hydrolysis by the recombinant Xyl2091 and Xyl2495. No activity against cellulose was observed for all enzymes. The presence of three xylanases ensures efficient xylan hydrolysis by M. roseus. The highly processive "free" endoxylanase Xyl2495 could hydrolyze xylan under moderate temperatures. Xylan hydrolysis at elevated temperatures could be accomplished by concerted action of two cell-bound xylanases; Xyl2090 that probably degrades xylans to long xylo-oligosaccharides, and Xyl2091 hydrolyzing them to xylose and xylobiose. The new endoxylanases could be useful for saccharification of lignocellulosic biomass in biofuels production, bleaching of paper pulp, and obtaining low molecular weight xylooligosaccharides.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.23-32
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• 1983

In the present study, two kinds of D-xylanases (1, 4-$\beta$-D-xylan xylanohydrolase (EC 3.2.1.8) were purified and characterized from crude extract of Aspergillus niger KG79. Xylanase I was most active at pH 5.0, whereas xylanse II at pH 4.0 Both enzymes demonstrated their maximum activity at 45$^{\circ}C$. They were relatively stable between pH 4.0 and 6.0 at 3$0^{\circ}C$ for 6 hours. Molecular weight of xylanse I and II were 12, 500 and 11, 500, respectively. Michaelis-Menten constants of xylanse I and II were 0.28% and 0.26% of xylan, respectively. Both enzymes could degrade commercial D-xylan to xylose, xylobiose, and xylotriose to the degree of about 10% of total reducing power. Xylanse I could, however, liberate arabinose from barley straw xylan in addition to xylose and xylooligasaccharides more rapidly than xylanase II. The degree of hydrolysis was about 25%. The reconstituted D-xylanase system with purified xylanases and $\beta$-xylosidase degraded commercial xylan and barley straw xylan to the degree of 28% and 54% respectively. The limit of hydrolysis by the enzymes was suggested to be resulted from the physical structure of the substrate.

• The Korean Journal of Food And Nutrition
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• v.10 no.3
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• pp.344-349
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• 1997

A strain of Bacillus sp. DSNC 101, isolated from soil, produced up to 305.0 units/ml of xylanase when grown on te medium containing 2.0% xylan, 2.0% yeast extract and 0.4% K2HPO4. The strain produced xylanase in the presence of xylan, soluble starch, rice straw, Avicel, maltose, and lactose as a sole carbon source, but the enzyme was not synthesized in the presence of xylose, glucose or arabinose. The crude xylanase preparation did not show hydrolytic activity towards cellulosic substrates and PNPX, a chromogenic substrate for $\beta$-xylosidase. The temperature and pH optima for the xylanase production were 4$0^{\circ}C$ and 8.0, respectively. Xylanase synthesis was repressed by glucose, but not by xylose. The hydrolysis products of xylan catalyzed with the culture filtrate were xylooligosaccharides such as xylobiose and xylotriose but xylose was not detected by tin layer chromatography.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.63-71
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• 1993

A xylanase (xylanase I) was purified 11.9-fold from the culture filtrate of Trichoderma koningii ATCC 26113 by the column chromatography on Sephadex G-75, SP-Sephadex C-50, DEAE-Sephadex A-50 and Sephadex G-50 with an overall yield of 8.2%. The molecular mass determined by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis was found to be a monomeric polypeptide of ca. 35 kDa. The isoelectric point of the enzyme was estimated to be 9.3. The optimal reaction pH and temperature are 5.8 and 55.deg.C, respectively. The enzyme is stable up to 60.deg.C, while 78% of its activity is lost after the incubation for 10 min at 70.deg.C. The enzyme hydrolyzes sylan with relatively high activity, as well as carboxymethyl cellulose and avicel. The $K_{m}$ values of the enzyme for oat-spelf sylan, larchwood xylan and Avicel were 3.5, 1.6 and 10. 1 mg/ml, respectively. The enzyme hydrolyzed oat-spelt sylan to sylose, sylobiose, sylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose, xylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose and xylotriose as the major products. The hydrolysis patterns indicate that xylanase I is endo-enzyme.e.