• 한국미생물·생명공학회지
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• 제31권2호
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• pp.157-164
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• 2003

알칼리성 xylanase를 생산하는 균주를 토양으로부터 분리한 후 동정을 실시한 결과 Bacillus alcaiophilus으로 판명되었다. B. alcalophilus AX2000으로 명명한 본 균주는 pH 10.5에서 생육이 가장 좋았으며 효소활성도 가장 높았고 배지 중에 탄소원과 질소원으로서 0.5%(w/v) birchwood xylan과 0.5%(w/v) polypeptone/yeast extract를 각각 사용하였을 경우에 최대의 xylanase생산성을 나타내었다. Xylanase의 생합성근 glucose에 의한 catabolite repression을 받았으며 고농도의 xylose에 의해 효소의 생합성이 저해되었다. 조효소의 최적활성은 pH 10과 $50^{\circ}C$에서 나타났으며, pH 5에서 11까지의 넓은 pH범위에서 활성이 안정하게 유지되었고 효소의 열 안정성은 20~$60^{\circ}C$에서 30분간 처리시 90%이상의 잔존활성을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제22권10호
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• pp.1388-1394
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• 2012

A xylanase-producing thermophilic strain, Geobacillus sp. TC-W7, was isolated from a hot spring in Yongtai (Fuzhou, China). Subsequently, the xylanase gene that encoded 407 amino acids was cloned and expressed. The recombinant xylanase was purified by GST affinity chromatography and exhibited maximum activity at $75^{\circ}C$ and a pH of 8.2. The enzyme was active up to $95^{\circ}C$ and showed activity over a wide pH range of 5.2 to 10.2. Additionally, the recombinant xylanase showed high thermostability and pH stability. More than 85% of the enzyme's activity was retained after incubation at $70^{\circ}C$ for 90 min at a pH of 8.2. The activity of the recombinant xylanase was enhanced by treatment with 10 mM enzyme inhibitors (DDT, Tween-20, 2-Me, or TritonX-100) and was inhibited by EDTA or PMSF. Its functionality was stable in the presence of $Li^+$, $Na^+$, and $K^+$, but inhibited by $Hg^{2+}$, $Ni^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Pb^{2+}$, $Fe^{3+}$, and $Al^{3+}$. The functionality of the crude xylanase had similar properties to the recombinant xylanase except for when it was treated with $Al^{2+}$ or $Fe^{2+}$. The enzyme might be a promising candidate for various industrial applications such as the biofuel, food, and paper and pulp industries.

• Asian-Australasian Journal of Animal Sciences
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• 제18권1호
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• pp.66-74
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• 2005

Three experiments were conducted to study the property of xylanase and the effects of xylanase in wheat-based diets on growth performance of broilers, respectively. Experiment 1 was performed in vitro to evaluate the effect of different pH and temperature on xylanase activity, and to evaluate the enzymic stability under different conditions. The results indicated that the optimum temperature and pH for xylanase activity were $50^{\circ}C$ and 4.5, respectively. The activity of enzyme solution was reduced rapidly after the treatment of water bath above $60^{\circ}C$ for 10 min. The enzyme was relatively stable at pH 3.5 to 8.0 and deteriorated when incubated at pH below 3.5. In Experiment 2, a total of 378 d-old male Arbor Acres broilers were randomly distributed to 7 different treatments with 6 replicates (9 birds) in each treatment. The treatments were as follows: (1) corn based diet (CS), (2) wheat based diet (WS), (3) WS+ 0.05% xylanase, (4) WS+0.15% xylanase, (5) WS+0.25% xylanase, (6) WS+0.35% xylanase, (7) WS+0.45% xylanase. The results showed that the body weight and feed/gain ratio of the broilers fed wheat-based diets have been significantly improved (p<0.05) compared to that fed corn-based diet in the first 3 wk. With regard to the wheat-based diets, the xylanase supplementation had a tendency to improve the growth performance in first 3 wk. After 3 wk, no significant difference (p>0.05) was found among all these different treatments. The supplementation of xylanase and the type of diets did not affect the feed intake but increased the concentration of triglyceride in serum. In Experiment 3, a total of 360 d-old male Arbor Acres broilers were assigned to 30 groups with 12 birds in each group randomly. These groups were then randomly distributed to 5 different treatments with 6 replicates within each treatment. The broilers of each treatment were fed one of the diets as follows: (1) Corn based diet, (2) White wheat based diet (WW) (3) White wheat based diet+0.25% xylanase, (4) Red wheat based diet, (5) Red wheat based diet+0.25% xylanase. The results showed that the body weight and feed/gain ratio had been significantly improved (p<0.05) by xylanase supplementation in the first 2 or 3 wk. The effect of xylanase in red wheat diet is a little higher than that used in white wheat diet. From the results of the present experiments, it can be concluded that the supplementation of Aspergillar xylanase can improve the performance of the broilers fed the wheat-based diet.

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.378-387
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• 1998

The DNA sequence immediately following the xynA gene of Bacillus stearothermophilus 236 ［about l-kb region downstream from the translational termination codon (TAA) of the xynA gene］was found to have an ability to enhance the xylanase activity of the upstream xynA gene. An 849-bp ORF was identified in the downstream region, and the ORF was confirmed to encode a novel protein of 283 amino acids designated as XaiF (xylanase-activity-increasing factor). From the nucleotide sequence of the xaiF gene, the molecular mass and pI of XaiF were deduced to be 32,006 Da and 4.46, respectively. XaiF was overproduced in the E. coli cells from the cloned xaiF gene by using the T7 expression system. The transcriptional initiation site was determined by primer extension analysis and the putative promoter and ribosome binding regions were also identified. Blast search showed that the xaiF and its protein product had no homology with any gene nor any protein reported so far. Also, in B. subtilis, the xaiF trans-activated the xylanase activity at the same rate as in E. coli. In contrast, xaiF had no activating effect on the co-expressed ${\beta}-xylosidase$ of the xylA gene derived from the same strain of B. stearothermophilus. In addition, the intracellular and extracellular fractions from the E. coli cells carrying the plasmid-borne xaiF gene did not increase the isolated xylanase activity, indicating that the protein-protein interaction between XynA and XaiF was not a causative event for the xylanase activating effect of the xaiF gene.

• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.397-404
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• 2013

Paenibacillus xylanilyticus KJ-03 was isolated from soil samples obtained from a field with Amorphophallus konjac plants. A gene encoding xylanase was isolated from KJ-03 and cloned using a fosmid library. The xynA gene encodes xylanase; it consists of 1,035 bp and encodes 345 amino acids. The amino acid sequence deduced from the P. xylanilyticus KJ-03 xylanase showed 81% and 69% identities with those deduced from the P. polymyxa E681 and Paenibacillus sp. HPL-001 xylanases, respectively. The xynA gene comprises a single domain, consisting of a catalytic domain of the glycosyl hydrolase (GH) 10 family. The xynA gene was expressed in Escherichia coli BL21 (trxB), and the recombinant xylanase was purified by Niaffinity chromatography. The purified xylanase showed optimum activity with birchwood xylan as a substrate at $40^{\circ}C$ and pH 7.4. Treatment with $Mg^{2+}$ and $Li^+$ showed a slight decrease in XynA activity; however, treatment with 5 mM $Cu^{2+}$ completely inhibited its activity. The results of the thin layer chromatography analysis indicated that the major hydrolysis product was xylobiose and small amounts of xylose and xylotriose. XynA showed increased activity with oat spelt xylan and birchwood xylan, but showed only slight activity with locust bean gum.

• 한국미생물·생명공학회지
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• 제20권5호
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• pp.559-564
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• 1992

섬유소 자원의 효율적 이용을 위하여 강력한 xylan 분해효소 생산 세균을 분리 및 선발하였으며, 형태학적, 배양학적 생리학적 특성을 확인한 결과 Bacillus sp. N25로 동정하였다. 이 균주에 의해서 생산된 효소를 ammonium sulfate precipitation, DAEA-sephadex A-50, Sephadex G-100 column으로 부분 정제하여 얻은 효소의 열안정성은 $50^{\circ}C$에서 30분간 처리시 80의 역가가 잔존 했으며, pH 안정은 pH 6-8 범위에서 30'C에서 10시간 방치 후에도 안정했으며, cellulose 분해능력은 없었으며, $Hg^{2+}$, $Ag^{2+}$, $Mn^{2+}$에 의해 저해되었다. 그리고 최종분해산물은 주로 xylose이므로 exo-type xylanase로 추정된다.

• 한국균학회지
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• 제25권4호통권83호
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• pp.297-304
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• 1997

흰목이균, 공생균, 혼합균(흰목이균과 공생균)의 xylanase 생산 최적 탄소원은 xylose이었고 흰목이균, 공생균의 질소원은 $KNO_3$, 혼합균의 질소원은 $(NH_4)_2SO_4$이었다. Xylanase 생산 최적 배양일수는 흰목이균과 혼합균은 5일, 공생균은 6일이었으며 최적온도는 흰목이균과 공생균이 $40^{\circ}C$, 혼합균이 $45^{\circ}C$이었다. Xylanase 생산 최적 pH는 흰목이균과 공생균이 pH 6.0, 혼합균이 pH 7.0이었으며, 금속이온의 영향은 흰목이균에서는 $Hg^{2+}$, 혼합균에서는 $Pb^{2+}$를 제외하고는 금속이온이 저해적으로 나타났다. 흰목이균, 공생균, 혼합균의 xylanase 열안전성은 $50^{\circ}C$까지 80% 이상 안정하였으며, 기질(Birchwood) 특이성에 관해서는 Birchwood로부터의 xylan이 가장 활성이 높았다. 흰목이균, 공생균, 혼합균의 xylanase 기질농도는 20mg/ml, 반응시간은 45분까지 활성이 급속히 상승하다가 그후 매우 완만한 반응속도를 나타내므로서 이를 기초로 하여 Km값을 얻은 결과, 흰목이균은 $6.25{\times}10^{-5}\;M$, 혼합균은 $5.2{\times}10^{-2}\;M$, 공생균은 $5.6{\times}10^{-2}\;M$로 흰목이균, 혼합균, 공생균 순으로 효소 기질과의 친화력이 약하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권11호
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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.

• 펄프종이기술
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• 제35권1호
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• pp.34-40
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• 2003

This study was carried out to select the useful bacteria which secret extracellula enzymes for enzymatic deinking agent of old newspaper. CMCase, FPase and xylanase activities of the bacteria liquid culture were measured at optimal growth conditions. Clear zone test was checked on the solid culture. The results of this study were as follow: Eight strains of 28 bacteria isolated from a paper mill soil ground were shown strong CMCase and xylanase activity with the clear zone test. The optimal pH and temperature for culture growth were 6~8 and 26~$34^{\circ}C$, respectively and optimal culture period were less than 60 hours. Based on CMCase, FPase and xylanase activity, strain No. 18, 21, 22 and 28 which were relatively higher than the other strains, were selected for further enzymatic deinking research.

• 미생물학회지
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• 제48권2호
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• pp.141-146
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• 2012

Paenibacillus woosongensis의 유전체 부분 염기서열로부터 유추된 xylanase 유전자를 PCR 증폭하여 대장균에 클로닝하였다. Xylanase 유전자는 211 아미노산으로 구성된 단백질을 코드하며 633 뉴클레오티드로 이루어졌다. 아미노산 잔기배열을 분석한 결과 xylanase는 glycosyl hydrolase family 11에 속하며 Paenibacillus의 xylanase와 85-89% 상동성을 보였다. Xylanase 유전자를 T7 promoter로 과잉발현한 결과 그 발현량이 높지 않았으며, 균체내 외에서 모두 효소활성이 관찰되었다. 재조합 대장균의 균체파쇄상등액을 사용하여 효소 반응특성을 조사한 결과 pH 5.5와 $60^{\circ}C$에서 최대 반응활성을 보였다. 한편 xylanase의 기질로 xylan과 xylooligosaccharides를 사용하였을 때 xylose와 xylotriose가 주된 최종 반응산물로 관찰되었으며 xylobiose는 분해하지 못하였으나 이보다 중합도가 큰 xylooligosaccharides는 분해하였다.