• Title/Summary/Keyword: Xylanase

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Purification and Characterization of Two Thermostable Xylanases from Paenibacillus sp. DG-22

  • Lee, Yong-Eok;Lim, Pyung-Ok
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1014-1021
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    • 2004
  • Two thermostable xylanases, designated XynA and XynB, were purified to homogeneity from the culture supernatant of Paenibacillus sp. DG-22 by ion-exchange and gel-filtration chromatography. The molecular masses of xylanases A and B were 20 and 30 kDa, respectively, as determined by SDS-PAGE, and their isoelectric points were 9.1 and 8.9, respectively. Both enzymes had similar pH and temperature optima (pH 5.0-6.5 and $70^{\circ}C$), but their stability at various temperatures differed. Xylanase B was comparatively more stable than xylanase A at higher temperatures. Xylanases A and B differed in their $K_m$ and $V_{max}$ values. XynA had a $K_m$ of 2.0 mg/ml and a $V_{max}$ of 2,553 U/mg, whereas XynB had a K_m$ of 1.2 mg/ml and a $V_{max}$, of 754 U/mg. Both enzymes were endo-acting, as revealed by their hydrolysis product profiles on birchwood xylan, but showed different modes of action. Xylotriose was the major product of XynA activity, whereas XynB produced mainly xylobiose. These enzymes utilized small oligosaccharides such as xylotriose and xylotetraose as substrates, but did not hydrolyzed xylobiose. The amino terminal sequences of XynA and XynB were determined. Xylanase A showed high similarity with low molecular mass xylanases of family 11.

Thermostable Xylanase Encoded by xynA of Streptomyces thermocyaneoviolaceus: Cloning, Purification, Characterization and Production of Xylooligosaccharides

  • CHOI JUN-HO;LEE OH-SEUK;SHIN JAE-HO;KWAK YUN-YOUNG;KIM YOUNG-MOG;RHEE IN-KOO
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.57-63
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    • 2006
  • We have cloned a xylanase gene (xynA) from Streptomyces thermocyaneoviolaceus. The deduced amino acid sequences of the XynA, including the active site sequences of glycosyl hydrolase family 10, showed high sequence homology with several xylanases assigned in this category. The XynA was overexpressed under an IPTG inducible T7 promoter control in E. coli BLR(DE3). The overproduced enzymes were excreted into culture supernatants and periplasmic space. The purified XynA had an apparent molecular mass of near 54 kDa, which corresponds to the molecular mass calculated from its gene. The optimum pH and temperature of the purified XynA were determined to be 5.0 and $65^{\circ}C$, respectively. The XynA retained over $90\%$ its activity after the heat treatment at $65^{\circ}C$ for 30 min. The XynA was highly efficient in producing xylose (X1), xylobiose (X2), xylotriose (X3), and xylotetraose (X4) from xylan.

Isolation and Characterization of a Xylanolytic Bacterium, Bacillus sp. MX47 (Xylanase 생산균 Bacillus sp. MX47의 분리 및 동정)

  • Chi, Won-Jae;Park, Da Yeon;Park, Jae-Seon;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.419-423
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    • 2012
  • A xylanolytic bacterial strain, MX47, was isolated from rotting plant matter in soil. The strain was aerobic and gram positive, and grew between pH 6.0 and 11.0. Cells were susceptible to thiostrepton and chloramphenicol. The major fatty acids (>3%) comprised 64.55% of iso-$C_{15:0}$, 22.76% of anteiso-$C_{15:0}$, and 3.92% of iso-$C_{17:0}$. The G/C content of the DNA was 44.15 mol%. The predominant respiratory quinone was menaquinone 7 (MK-7). Searches for 16S rRNA gene sequence similarity as well as phylogenetic analyses strongly suggested that the strain should be classified to the genus Bacillus. However, its biochemical characteristics, including acid production and enzyme activities, are different from those of other Bacillus strains in the same clade, and therefore, we propose the name Bacillus sp. MX47.

Elemental Chlorine free Bleaching of Kraft Pulps with Enzymes( I )-Oakwood Kraft Pulp- (효소를 이용한 크라프트펄프의 무감소표백-신갈나무 크라프트펄프-)

  • 강진하;박성종;임현아
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.30 no.1
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    • pp.44-58
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    • 1998
  • This study was carried out to bleach the Oakwood kraft pulp without the elemental chlorine using the xylanase or wastewater(We : wastewater enzymes) effluented from the submerged biofilter reactor containing the fungi, Phanerorhaete sordida YK-624. So in this research, the proper treatment conditions (pH, temperature, dosage and time) were investigated respectively. And after the various kinds of multistage bleaching of pulps, the properties of pulps were tested. From the experimental results, we can conclude as follows. In the treatments of Oakwood kraft pulps with xylanase, the proper pH, temperature, enzyme dosage and time were 8.0, $35^{\circ}C$ , 400 EXU/kg and 1 hr. respectively. And in the case of treatment with a wastewater(We) effluented from the submerged biofilter reactor, the proper pH, temperature and time were 5.5, $37^{\circ}C$ and 2 hr. respectively. On the other hand, Oakwood kraft pulps were bleached by the method of a multistage bleaching using xylanase or We instead of elemental chlorine Consequently the strengthes and brightnesses of pulps bleached by the method mentioned above were lower than those of pulp bleached by the conventional method using the elemental chlorine. But it is possible to improve the brightnesses through the increase of chlorine dioxide dosage or use of hydrogen peroxide in the final bleaching stage.

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Production of Xylanolytic Enzyme Complex from Aspergillus flavus using Agricultural Wastes

  • Kim, Jeong-Dong
    • Mycobiology
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    • v.33 no.2
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    • pp.84-89
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    • 2005
  • Five types of agricultural wastes were used for the production of xylanolytic enzyme by Aspergillus flavus K-03. All wastes materials supported high levels of xylanase and ${\beta}-xylosidase$ production. A high level of proteolytic activity was observed in barley and rice bran cultures, while only a weak proteolytic activity was detected in corn cob, barley and rice straw cultures. Maximum production of xylanase was achieved in basal liquid medium containing rice barn as carbon source for 5 days of culture at pH 6.5 and $25^{\circ}C$. The xylanolytic enzyme of A. flavus K-03 showed low thermostability. The times required for 50% reduction of the initial enzyme activity were 90 min at $40^{\circ}C$, 13 min at $50^{\circ}C$, and 3 min at $60^{\circ}C$. Xylanolytic activity showed the highest level at pH $5.5{\sim}10.5$ and more than 70% of the original activity was retained at pH 6.5 and 7.0. The higher stability of xylanolytic enzymes in the broad range of alkaline pH is useful for utilization of the enzymes in industrial process requiring in alkaline conditions. Moreover, the highest production of xylanolytic enzyme was obtained when 0.5% of rice bran was supplied in basal liquid medium. SDS-PAGE analysis revealed a single xylanase band of approximately 28.5 kDa from the culture filtrates.

Screening of Microorganisms Secreted High Efficient Enzymes and Properties of Enzymatic Deinking for Old Newsprint(IV) -Culture conditions of fungal extracellular enzyme production for biological deinking system- (고효율 효소를 분비하는 균주의 선발 및 신문고지의 효소탈묵 특성(제4보) -고지탈묵용 Fungal Cellulase와 Xylanase의 생산-)

  • Park Seong-Cheol;Kang Jin-Ha;Lee Yang-Soo
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.37 no.1 s.109
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    • pp.38-46
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    • 2005
  • This study was focused on the optimum culture condition in CMCase, FPase and xylanase activities of two fungal strains that secret extracellular enzymes for using enzymatic deinking agent to old newsprint. The results of this study were as follows. When Fusarium pallidoroseum was grown on the medium, containing of rice bran+xylan $2.0\%,\;peptone\;0.6\%,\;KH_2PO_4\;0.075\%\;and\;MnSO_4\;0.06\%\;with\;pH\;9.0,\;at\;29^{\circ}C$ for 6 days, the quantitative degree of extracellular enzyme production was the highest. Optimum culture condition for Aspergillus niger was pH 5.0, $27^{\circ}C$ incubating temperature and 7 days incubation period on liquid medium, containing of CMC+xylan $2.5\%,\;yeast\;extract\;0.4\%,\;K_3PO_4\;0.05\%\;and\;CaCl_2+FeSO_4\;0.08\%$. Aspergillus niger was fairly higher FPase and xylanase activities than Trichoderma reesei ATCC 28217.

Elemental Chlorine Free Bleaching of Kraft Pulps with Enzymes(II) -Pinewood Kraft Pulp- (효소를 이용한 크라프트펄프의 무감소표백(제2보) -소나무 크라프트펄프-)

  • 강진하;박성종;정인수
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.30 no.3
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    • pp.84-96
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    • 1998
  • This study was carried out to bleach the Pinewood kraft pulp without the elemental chlorine using the xylanase or wastewater(We:wastewater enzymes) effluented from the submerged biofilter reactor containing the fungi, Phanerochaete sordida YK-624. So in this research, the proper treatment conditions(pH, temperature, dosage and time) were investigated respectively. And after the various kinds of multistage bleaching of pulps, the properties of pulps were tested. From the experimental results, we can conclude as follows. In the treatments of Pinewood kraft pulps with xylanase, the proper pH, temperature, enzyme dosage and time were 8.0, $35^{\circ}C, 400EXU/kg and 3 hr. respectively. And in the case of treatment with a wastewater(We) effluented from the submerged biofilter reactor, the proper pH, temperature and time were 5.0, $37^{\circ}C and 3 hr. respectively. On the other hand, Pinewood kraft pulps were bleached by the method of a multistage bleaching using xylanase or We instead of elemental chlorine. Consequently, the strengthes and brightnesses of pulps bleached by the method mentioned above were lower than those of pulp bleached by the conventional method using the elemental chlorine. But it is possible to improve the brightnesses through the increase of chlorine dioxide dosage or use of hydrogen peroxide in the final bleaching stage.

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Hydrolysis of Agricultural Residues and Kraft Pulps by Xylanolytic Enzymes from Alkaliphilic Bacillus sp. Strain BK

  • Kaewintajuk Kusuma;Chon Gil-Hyong;Lee Jin-Sang;Kongkiattikajorn Jirasak;Ratanakhanokchai Khanok;Kyu Khin Lay;Lee John-Hwa;Roh Min-Suk;Choi Yun-Young;Park Hyun;Lee Yun-Sik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1255-1261
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    • 2006
  • An alkaliphilic bacterium, Bacillus sp. strain BK, was found to produce extracellular cellulase-free xylanolytic enzymes with xylan-binding activity. Since the pellet-bound xylanase is eluted with 2% TEA from the pellet of the culture, they contain a xylan-binding region that is stronger than the xylan-binding xylanase of the extracellular enzyme. The xylanases had a different molecular weight and xylan-binding ability. The enzyme activity of xylanase in the extracellular fraction was 6 times higher than in the pellet-bound enzyme. Among the enzymes, xylanase had the highest enzyme activity. When Bacillus sp. strain BK was grown in pH 10.5 alkaline medium containing xylan as the sole carbon source, the bacterium produced xylanase, arabinofuranosidase, acetyl esterase, and $\beta$-xylosidase with specific activities of 1.23, 0.11, 0.06, and 0.04 unit per mg of protein, respectively. However, there was no cellulase activity detected in the crude enzyme preparation. The hydrolysis of agricultural residues and kraft pulps by the xylanolytic enzymes was examined at 50$^{\circ}C$ and pH 7.0. The rate of xylan hydrolysis in com hull was higher than those of sugarcane bagasse, rice straw, com cop, rice husk, and rice bran. In contrast, the rate of xylan hydrolysis in sugarcane pulp was 2.01 and 3.52 times higher than those of eucalyptus and pine pulp, respectively. In conclusion, this enzyme can be used to hydrolyze xylan in agricultural residues and kraft pulps to breach and regenerate paper from recycled environmental resources.

The High Production of Cellulolytic Enzymes using Cellulosic Wastes by a Fungus, strain FJ1. (섬유소폐기물을 이용한 사상균 FJ1의 섬유소 분해효소의 고생산)

  • 유승수;김경철;오영아;정선용;김성준
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.172-176
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    • 2002
  • A filamentous microorganism, strain FJ1, was isolated from completely rotten wood for the production of cellulolytic enzymes. For the production of the enzymes, cellulolsic wastes were used as carbon sources of strain FJ1 and rice straw showed higher enzyme activities than sawdust and pulp. The activities of CMCase, xylanase, $\beta$-glucosidase, and avicelase were 2.95, 5.89, 0.45, and 0.12 unit/ml by use of rice straw, respectively. To enhance production of the enzymes, the mixture substrate of rice straw and cellulosic materials were investigated as carbon sources. The highest activities of CMCase, $\beta$-glucosidase, and avicelase were found in the mixture of rice straw (0.5%, w/v) and avicel (0.5%, w/v), and the highest xylanase was obtained at the mixture ratio of 0.71%(w/v) and 0.29%(w/v). Addition of 0.1%(w/v) peptone showed enhanced production of the cellulolytic enzymes in which the activities of CMCase, xylanase, $\beta$-glucosidase, and avicelase were 19.23, 27.18, 1.28, and 0.53 unit/ml, respectively. The production of the enzymes using rice straw was efficiently induced in the presence of avicel and pulp containing cellulose. In particular, a medium composed of rice straw (0.5%, w/v) and pulp (0.5%, w/v) yielded larger cellulolytic enzymes: CMCase 24.3 unit/ml, xylanase 38.7 unit/ml, $\beta$-glucosidase 1.5 unit/ml, and avicelase 0.6 unit/ml. The filamentous microorganism, strain FJ1 utilized various cellulosic wastes as carbon sources and will be expected as a favorable candidate for biological saccharification of cellulosic wastes.

Effects of Low Level Water-soluble Pentosans, Alkaline-extractable Pentosans, and Xylanase on the Growth and Development of Broiler Chicks

  • Sheng, Q.K.;Yang, L.Q.;Zhao, H.B.;Wang, X.L.;Wang, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.9
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    • pp.1313-1319
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    • 2013
  • This study investigated the effects of low levels of water-soluble pentosans (WSP), alkaline-extractable pentosans (AEP), and xylanase on the growth and organ development of broiler chicks. Three hundred and fifty 1-d-old female broiler chicks were randomly allocated into seven experimental groups of five pen replicates, with ten chicks per replicate. The control group consumed a corn-soybean meal-based diet. Six dietary treatment groups consumed the basal diet supplemented with one of the following: WSP at 50 mg/kg (WSP50) or 100 mg/kg (WSP100); AEP at 50 mg/kg (AEP50) or 100 mg/kg (AEP100); or xylanase at 3 mg/kg (Xase3) or 6 mg/kg (Xase6). Data including the body weight, digestive organ weights, gut length, rectal digesta viscosity, and gut microflora and pH were collected on d 5, 10, and 15. When compared to the control group, WSP50 promoted body weight gain and organ growth throughout the study, calculated as 3-d averages (p<0.05). WSP100 increased weight gain and enhanced organ development (proventriculus, gizzard, and gut) on d 10 (p<0.05), but the 3-d averages were not different from the control group except for the weight of gizzard. Both Xase3 and Xase6 increased the 3-d average weight gain and the growth of the gizzard (p<0.05). WSP50 increased the digesta viscosity compared to Xase3 on d 10 and 15 (p<0.05). WSP50, Xase3, and Xase6 increased the concentration of Lactobacillus in the rectum when compared to the control group (p<0.05), but only Xase3 lowered the digesta pH in the ileum and cecum on d 10 and 15. AEP had minimal influence on the growth and organ development of broilers. The results showed that low levels of WSP, AEP, and xylanase had different effects and underlying mechanisms on the growth and organ development of broiler chicks. WSP50 could increase the growth performance of broilers fed a corn-soybean meal-based diet.