• Asian-Australasian Journal of Animal Sciences
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• 제30권1호
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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.

• 한국미생물·생명공학회지
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• 제44권3호
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• pp.370-375
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• 2016

가정식 된장에서 mannanase 생산균으로 분리된 Bacillus sp. YB-1401와 B. amyloliquefaciens YB-1402로부터 2개의 xylanase 유전자가 대장균으로 클로닝 되었으며 그 염기서열이 결정되었다. 두 xylanase 유전자는 213개 아미노산 잔기의 단백질을 코드하는 642 nucleotides로 동일하게 구성되었다. Xyn1401과 Xyn1402로 명명된 YB-1401과 YB-1402 xylanase의 아미노산 배열은 127번째 잔기인 Asn과 Lys을 제외하고는 모두 동일하였고, glycosyl hydrolase family 11에 속하는 xylanase의 아미노산 배열과 상동성이 높았다. 두 효소의 signal peptide는 SignalP4.1 프로그램에 의해 아미노 말단의 28 잔기 배열로 동일하게 예측되었다. 두 효소는 91−94%가 재조합 대장균의 배양상등액에 존재하였으므로 대장균에서 효과적으로 분비되는 것으로 판단되었다. Xyn1401과 Xyn1402의 최적 반응조건은 50℃와 pH 6.0, 55℃와 pH 6.5로 각각 달랐으며 이로 보아 오직 한 개의 아미노 잔기의 차이가 온도와 pH에 따른 효소 활성에 영향을 미치는 것으로 확인되었다. 또한 두 효소는 열안정성도 서로 약간 차이가 있었다.

• 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.

• 한국식품영양과학회지
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• 제46권9호
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• pp.1114-1121
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• 2017

전통장류로부터 식품 유해요소를 생성하지 않는 Bacillus 균주를 분리하여 세포외효소 활성(amylase, protease, cellulase, xylanase)을 측정한 후, 단백질 분해 활성이 우수한 14개 균주와 비교균주 1균주를 선발하였다. 선발된 균주에 대해 16S rRNA 유전자를 이용한 균주 동정을 실시한 결과, B. amyloliquefaciens 10종, B. methylotrophicus 1종, B. velezensis 1종, B. subtilis 3종이 분리 동정되었다. 그중 B. subtilis JBG17019, B. amyloliquefaciens JBD17076, B. amyloliquefaciens JBD17109 균주에서 식중독미생물에 대한 증식 억제능이 확인되었다. Glutamic acid 대사와 관련한 발효특성을 확인하기 위하여 선발된 Bacillus 균주에 대해 glutamate, glutamine 및 ${\gamma}$-PGA 생성능을 측정하였다. 발효특성과 ${\gamma}$-PGA 생성능에 대한 다변량 요인분석을 주성분(PCA) 추출법으로 분석한 결과, PC1(효소 활성(amylase, cellulase, xylanase), PC2(${\gamma}$-PGA 생성능) 및 PC3(protease, glutamate 및 glutamine)의 3가지 주성분이 분류되었다. 주성분(PC)의 추출에 따라 B. amyloliquefaciens JBD17076 및 B. subtilis JBG17019 균주는 우수한 효소 활성 및 ${\gamma}$-PGA 생성을 하는 것으로 평가되었다.

• 펄프종이기술
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• 제37권1호
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• pp.25-37
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• 2005

This study was carried out to examine the optimal cultural condition in enzyme activities of CMCase, FPase and xylanase in selected strains which secret extracellular enzymes for using deinking agent to old newsprint. The results of this study were as follow: The production of enzyme by Bacillus pumilus I was maximal as grown on the medium, containing of rice bran+xylan $2.0\%$, peptone $0.8\%,\;K_2HPO_4\;0.1\%\;and\;CaCl_2\;0.06\%$ at pH 8.0 and $28^{\circ}C$ for 72 hours. Optimal cultural condition of B. subtilis I was avicel+xylan $3.5\%,\;urea\;0.4\%,\;K_3PO_4\;0.1\%\;and\;CaCl_2\;0.015\%$ at pH 9.0 and $28^{\circ}C$ for 36 hours. The maximal enzyme production was observed in the medium, containing of avicel+xylan $3.5\%,\;urea\;1.6\%\;and\; K_2HPO_4\;0.125\%$ with pH 9.0 when B. pumilus II was cultured at $28^{\circ}C$ for 60 hours. The production of enzyme by B. subtilis IT was maximal as grown on the medium, containing of xylan $2.0\%,\;yeast\; extract\;0.6\%,\;K_2HPO_4\;0.1\%\;and\;ZnSO_4\;0.04\%$ at pH 8.0 and $34^{\circ}C$ for 36 hours. The activities of FPase and xylanase in tested 4 strains were not much different with Thermomonospora fusca.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1471-1480
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• 2010

An extremely thermostable xylanase gene, xynB, from the hyperthermophilic bacterium Thermotoga maritima MSB8 was successful expressed in Kluyveromyces lactis. The response surface methodology (RSM) was also applied to optimize the medium components for the production of XynB secreted by the recombinant K. lactis. The secretion level (102 mg/l) and enzyme activity (49 U/ml) of XynB in the optimized medium (yeast extract, lactose, and urea; YLU) were much higher than those (56 mg/l, 16 U/ml) in the original medium (yeast extract, lactose, and peptone; YLP). The secretory efficiency of mature XynB was also improved when using the YLU medium. When the mRNA levels of 13 characterized secretion-related genes in the K. lactis cultured in YLP and YLU were detected using a semiquantitative RT-PCR method, the unfolded protein response (UPR)-related genes, including ero1, hac1, and kar2, were found to be up-regulated in the K. lactis cultured in YLU. Therefore, the nutrient ingredients, especially the nitrogen source, were shown to have a significant influence on the XynB secretory efficiency of the host K. lactis.

• 한국버섯학회지
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• 제13권4호
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• pp.305-309
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• 2015

Cellulase와 xylanase 분비능이 우수한 부숙촉진 세균을 분리하기 위하여 진주시 집현면 소재의 느타리버섯 재배농장으로부터 느타리버섯 수확후배지를 수집하였다. 느타리버섯 수확후배지로부터 19종의 균주를 분리하였으며 이 중 cellulase와 xylanase을 동시에 분비하는 균주를 최종 선발하여 CA105로 명명하였다. Bacillus ID kit와 VITEK 2 system를 이용하여 분리균 CA105의 생화학적 특성을 조사한 결과에서도 분리균 CA105은 B. subtilis와 유사한 특징을 나타내었으며 16S rRNA 염기서열 분석 결과에서는 B. subtilis와 98.9%의 상동성을 나타내었다. 이와 같은 결과를 종합하여 분리균 CA105은 Bacillus subtilis CA105로 동정되었다. 분리균이 분비하는 cellulase와 xylanase 활성은 분리균이 증식함에 따라 대수증식기 중반부터 급격히 증가하였고 정지기에 진입하면 효소활성이 더 이상 증가하지 않는 것으로 나타났다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.477-480
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• 2002

A thermostable xylanase from Thermotoga maritima (Xyn B) cleaves several pNP-glycosides of monosaccharides. We found that the initial product of the cleavage of pNP-xyloside (pNP-Xy1) was a disaccharide, not xylose, indicating that xylosyl unit of pNP-Xyl was transglycosylated to another pNP-Xyl. We determined that the disaccharide was xylobiose which has the linkage of the ${\beta}$ 1-4, and described the reaction mechanism of the Xyn B. Also, we produced the several pNP-glycosides and propyl-disaccharides from the transglycosylation of Xyn B with varial glycosides and/or 1-propanol. All reaction products were purified by column chromatography (Toyo-pearl HW-40C, 45 cm${\times}$2.5 cm or 45 cm ${\times}$ 2.5 cm${\times}$ 2). The isolated products were analyzed by means of 1D and 2D NMR.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.534-538
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• 2001

The xynA gene encoding an alikali-tolerant endo-1,4-${\beta}$-xylanase (XYN) was cloned from the alkalophilic Bacillus pumilus A-30. The nucleotide sequence of a 974-bp DNA fragment containing the xynA was determined. An ORF of 684 nucleotides that encoded a protein of 228 amino aicds was detected. Asparagine-71 of XYN from B. Pumilus A-30 showed to be highly conservative in alkaline xylanases of family G/11, upon comparing the amino acid sequences of 17 family G/11 xylanases. Site-directed mutation of N71D of the xynA gene resulted in a decrease of 12.4% in the specific acitivity and a significant decline in the enzyme activity in the alkaline pH range.

• Journal of Microbiology and Biotechnology
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• 제22권8호
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• pp.1084-1091
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• 2012

Xylooligosaccharides are functional foods mainly produced during the hydrolysis of xylan by physical, chemical, or enzymatic methods. In this study, production of xylobiose was investigated using oil palm empty fruit bunch fiber (OPEFB) as a source material, by chemical and enzymatic methods. Xylanase-specific xylan hydrolysis followed by xylobiose production was observed. Among different xylanases, xylanase from FXY-1 released maximum xylobiose from pretreated OPEFB fiber, and this fungal strain was identified as Aspergillus terreus and subsequently deposited under the accession Number MTCC- 8661. The imperative role of lignin on xylooligosaccharides enzymatic synthesis was exemplified with the notice of xylobiose production only with delignified material. A maximum 262 mg of xylobiose was produced from 1.0 g of pretreated OPEFB fiber using FXY-1 xylanase (6,200 U/ml) at pH 6.0 and $45^{\circ}C$. At optimized environment, the yield of xylobiose was improved to 78.67 g/100 g (based on xylan in the pretreated OPEFB fiber).