• 한국미생물·생명공학회지
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• 제51권1호
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• pp.59-68
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• 2023

Xylanase is an industrially relevant enzyme used for the production of xylobiose and xylose. Various methods are used to enhance the microbial yield of xylanase. In the present study, co-culturing of Bacillus subtilis and Bacillus pumilus were investigated using submerged fermentation for xylanase production, which was markedly increased when sal, sagwan, newspaper, wheat bran, and xylan were used as single carbon sources. Maximum xylanase production was reported after 5 days of incubation in optimized media at pH 7.0 and 37℃, resulting in 2.69 ± 0.25 µmol/min by coculture. The 1:1 ratio of sal and sagwan in optimized production media was shown to be suitable for xylanase synthesis in submerged fermentation (SMF). In comparison to mono-culture using B. pumilus and B. subtilis, co-culturing resulted in an overall 3.8-fold and 2.15-fold increase in xylanase production, respectively.

• Journal of Animal Science and Technology
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• 제51권5호
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• pp.427-432
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• 2009

본 연구에서는 xylanase, mannanase를 생산하는 미생물균주를 분리하기 위해 1차로 토양 및 낙엽 등에서 곰팡이 균주를 약 50여주 분리하였으며, 1차 선발된 균주로부터 xylanase, mannanase를 생산하는 미생물 분리를 위해 xylanase 생성균주 선별배지, mannanase 생성균주 선별배지에 single colony를 접종한 후 clear zone이 생기는 15종의 균주를 2차 선발하였다. 2차 선발된 균주를 개별적으로 배양하여, DNS 방법을 활용하여 xylanase, mannanase 효소활성을 측정하여 6종의 균주를 선발하였다. 선별된 균주는 액상배양에서 생산한 xylanase, mannanase 효소활성이 각각 0.9~1.6 unit/mL, 0.2~0.4 unit/mL 범위로 나타났다. 이 중 결과가 좋은 3종을 선정하여 고상배양으로 배양한 균주의 xylanase, mannanase 효소활성은 각각 103.7~220.0 unit/g, 20.1~40.3 unit/g으로 분석되었다. 선별된 3종의 균주중 xylanase, mannanase 효소활성이 각각 197.3 unit/g, 39.9 unit/g으로 가장 높은 E-3 균주를 최종적으로 선발하였다. 최종으로 분리한 E-3 균주는 형태학적 특징과 DNA 염기서열을 비교한 결과 Aspergillus niger와 99% 일치하였다.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.414-419
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• 1996

The xylnX gene encoding a xylanase from Clostridium thermocellum ATCC27405 was cloned in the plasmid pJH27, an E. coli-Bacillus shuttle vector and the resultant recombinant plasmid, pJX18 was transformed into E. coli HB101. The overexpressed xylanase was found to be secreted into the periplasmic space of the recombinant E. coli cells. The crude enzyme was obtained by treating the E. coli cells with lysozyme, and purified by DEAE-Sepharose column chromatography. Molecular wieght of the xylanase was estimated to be 53 kDa by gel filtration. The pI value was determined to be pH 8.8. The N-terminal sequence of the enzyme protein was Asp-Asp-Asn-Asn-Ala-Asn-Leu-Val-Ser-Asn which was considered to be the sequence of that of the mature form protein. The Km value of the enzyme for oat spelt xylan was calculated to be 2.63 mg/ml and the Vmax value was $0.47 {\mu}mole/min$. The xylanase had a pH optimum for its activity at pH 5.4 and a temperature optimum at $60^{\circ}C$. The enzyme hydrolyzed xylan into xylooligosaccharides which were composed mainly of xylobiose (40%) and xyloltriose (12%) after 5 hour reaction. This result indicates that the xylanase from C. thermocellum ATCC27405 is an endo-acting enzyme.

• 한국균학회지
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• 제25권2호통권81호
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• pp.91-100
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• 1997

회분배양과 유가배양을 이용하여 호알칼리성 Cephalosporium sp. RYM-202로부터 alkaline carboxymethyl cellulase (CMCase)와 Xylanase의 생산을 위한 배양조건에 대하여 조사하였다. 조사한 탄소기질 중에서 밀기울이 두 효소의 생산에 가장 효율적이었다. 또한 CMCase는 carboxymethyl cellulose (CMC)를 탄소기질로 첨가한 배양액에서, 반면에 xylanase의 경우에는 xylan을 기질로 하였을 때 높은 생산량을 나타냄으로서 유도기질 특이성을 보였는 바, 이 결과는 Cephalosporium sp. RYM-202에서의 CMCaee와 xylanase의 생합성이 효소 유도 수준에서 독립적으로 조절됨을 시사해 준다. 조사된 질소원 중에서는 무기질소원인 $NaNO_3$가 효소생산에 효과적이었으며, 이들 효소의 최대 생산을 위한 배양온도와 pH는 각각 $20^{\circ}C$와 9.0인 것으로 나타났다. 한편, 발효조에서의 회분배양을 통한 효소생산의 경우, 밀기울의 농도를 5%까지 증가시킴에 따라 효소생산량은 증가되었으나 catabolite repression에 의해 효소생산의 지연과 생산력의 감소를 초래하였다. 이러한 문제점은 탄소원의 간헐적 공급에 의한 유가배양을 통해서 어느 정도 해결될 수 있는 것으로 나타났으며, 밀기울의 최종농도가 5% 되게 공급된 유가배양 시 CMCase와 xylanase의 최대 효소생산량은 각각 0.39 및 9.2 units/ml 이었으며, 이는 같은 농도의 밀기울을 함유하는 회분배양 시 획득된 효소활성에 비해 각각 1.22배와 1.36배 증가된 것이다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권7호
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• pp.945-951
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• 2013

In vitro studies were undertaken to develop an appropriate fibrolytic enzymes cocktail comprising of cellulase, xylanase and ${\beta}$-D-glucanase for maize stover with an aim to increase its nutrient utilization in sheep. Cellulase and xylanase added individually to ground maize stover at an increasing dose rates (0, 100, 200, 400, 800, 1,600, 3,200, 6,400, 12,800, 25,600, 32,000, 38,400, and 44,800 IU/g DM), increased (p<0.01) the in vitro dry matter digestibility and in vitro sugar release. The doses selected for studying the combination effect of enzymes were 6,400 to 32,000 IU/g of cellulase and 12,800 to 44,800 IU/g of xylanase. At cellulase concentration of 6,400 IU/g, IVDMD % was higher (p<0.01) at higher xylanase doses (25,600 to 44,800 IU/g). While at cellulase doses (12,800 to 32,000 IU/g), IVDMD % was higher at lower xylanase doses (12,800 and 25,600 IU/g) compared to higher xylanase doses (32,000 to 44,800 IU/g). At cellulase concentration of the 6,400 to 32,000 IU/g, the amount of sugar released increased (p<0.01) with increasing levels of xylanase concentrations except for the concentration of 44,800 IU/g. No effect of ${\beta}$-D-glucanase (100 to 300 IU/g) was observed at lower cellulase-xylanase dose (cellulase-xylanase 12,800 to 12,800 IU/g). Based on the IVDMD, the enzyme combination cellulase-xylanase 12,800 to 12,800 IU/g was selected to study its effect on feed intake and rumen fermentation pattern, conducted on 12 rams (6 to 8 months; $20.34{\pm}2.369$ kg body weight) fed 50% maize stover based TMR. The total volatile fatty acids (p<0.01) and ammonia-N concentration was higher in enzyme supplemented group, while no effect was observed on dry matter intake, ruminal pH and total nitrogen concentration.

• 한국미생물·생명공학회지
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• 제20권5호
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• pp.543-550
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• 1992

Hemicellulose를 효소에 의하여 분해기키기 위하여 Aspergillus niger NRC 107로부터 xylanase와 $\beta$-xylosidase의 생산조건에 대하여 조사하였다. 이들 효소 생산 최적 pH는 6.0이었으며 탄소원 중 corn-cob xylan과 질소원 중 $NaNO_{3}$이 효소 생산에 제일 좋은 기질이었으며 이들의 최적농도는 각각 15g/l와 2.67g/l이다. 이들 효소는 인산염($KH_2P0_4$)과 Tween-80의 농도를 조절하므로 수율을 높일 수 있으며 wheat bran은 xylanase의 생산에L(-) sorbose는 $\beta$-xylosidase의 생산에 좋은 영향을 주었다. 이와 같이 이 Aspergillus niger NRC 107의 회분식 배양에서 이들 효소에 영향을 미치는 생산조건을 최적화한 결과 xylanase는 39.43 units/ml, $\beta$-xylosidase는 4.2units/ml 까지 생산할 있었다.

• 한국미생물·생명공학회지
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• 제17권2호
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• pp.85-87
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• 1989

토양으로부터 분리한 Xylose 발효효모 X-6-41 (Candida sp.)의 xylanase 효소생성 조건을 조사해 본 결과, 본 실험에 사용한 X-6-41 균주는 배지조성중 yeast extract 혹은 polypeptone이 첨가되면 xylanase 효소생성이 없었다. 즉 특정 아미노산인 isoleucine에 의해서는 그 생성이 억제되었고, tryptophan 25$\mu$g/$m\ell$ 농도의 존재하에서는 xylanase 생성이 증가했다.

• 한국미생물·생명공학회지
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• 제17권2호
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• pp.88-93
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• 1989

Xylan성 biomass로부터 직접 alcohol을 얻고자 xylose 발효효모 X-6-41 균주의 NTG mutant인 X-6-41-1(his-) 균주와 xylanase 분비효모인 XB-33 의 NTG mutant XB-33-37(Arg-) 균주를 세포융합 시켰다. 원형질체 생성조건은 KYPX(XB-33), KYPD(X-6-41)에서 대수증식기 말기까지 증식한 세포를 집균하여 zymolyase(0.25mg/$m\ell$), cellulase (4mg/$m\ell$)와 100mM 2-mercaptoethanol 처리시 protoplast화 율은 X-6-41 경우 80%, XB-33인 45%로 나타났다. 선별된 융합체의 탄소자화능은 parent의 자화능을 서로 보완하였음을 확인하였고 xylan에서의 alcohol 발효는 2% xylan에서 발효 15 일만에 총당에 대해 0.28% alcohol 농도를 나타내었다. Parent와 fusant의 xylanase 활성은 완전배지 보다 최소배지상에서 높았는데 이것은 xylose 발효효모(X-6-41)의 경우 isoleucine에 의한 xylanase production inhibition 효과와 일치하였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.337-340
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• 2001

The production of xylanase from Aspergillus niger mutant in SSF was optimized by' using statistical experimental designs. An inoculum size of $5{\times}10^5$ spores/g. initial moisture content of 65 %. cultivation time of 5 days and 10 times concentration of basal medium containing 50 times concentration of CSL were optimum for xylanase production ‘ Under the optimized conditions. the activity and productivity of xytanase obtained after 5 days of fermentation were 5.071 IU/gram of rice straw and 14.790 IU/l.h. respectively.

• Journal of Applied Biological Chemistry
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• 제57권4호
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• pp.313-320
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• 2014

Black liquor (BL) is a by-product of rice straw pulping process. It is a low costs raw material for production value-adding proteins and enzymes, which has been paid more and more attention to reduce its environmental pollution. Mixed cultures of micelial fungi, Trichoderma reesei Northern Regional Research Laboratory (NRRL)11236, Trichoderma reesei NRRL 6165 and Aspergillus niger strains NRC 5A, NRC 7A, and NRC 9A were evaluated for their ability to produce xylanase using crude hemicellulose (CHC) prepared from BL and peat moss as an inert support under solid state fermentation (SSF). The most potent strains, A. niger NRC 9A (818.26 U/g CHC) and T. reesei NRRL 6165 ($100.9{\pm}57.14$ U/g CHC), were used in a mixed culture to enhance xylanase production by co-culturing under SSF. In the mixed culture, xylanase production ($1070.52{\pm}12.57$ U/g CHC) was nearly1.3 and 10.6-fold increases over the activities attained in their monocultures, A. niger NRC 9A and T. reesei NRRL 6165, respectively. Optimization of the culture parameters of the mixed culture SSF process, concentration of ammonium sulfate and corn steep liquor, CHC/peat moss ratio, inoculum size and ratios of the two strains, initial pH value, initial moisture content and incubation time, exhibited a significant increase ($2414.98{\pm}84.02$ U/g CHC) in xylanase production than before optimization.