• 한국식품영양학회지
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• 제6권4호
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• pp.314-321
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• 1993

We have cloned the Bacillus cellulyticus K-12 avicelase (Avi, E.C.3.2.1.4) gene (ace A) In E. coli. This was accompanied by using the vector PT7T3U 19 and Hind W -Hind m libraries of Bacillus cellulyticus K-12 chromosomal inserts created in 5.cofi. The Libraries were screened for the expression of avicelase by monitoring the immunoreaction of the anti-avicelase (immunoscreening). Positive clones (Ac-3, Ac-5, and Ac-7) contained the identical 3.5kb Hind III fragment as determined by restriction mapping and Southern hybridization, and expressed avicelase efficiently and constituvely using its own promoter in the heterologous host. From the immunoblotting analysis, a polypeptide which showed a CMCase activity with an Mr of 54000 was detected.

• 한국미생물·생명공학회지
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• 제20권2호
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• pp.164-168
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• 1992

Cellulomonas sp.YE-5가 생산하는 cellulase를 분리, 정제하여 효소의 특성을 알아보았다. Avicelase, CMCase, $\beta$-glucosidase의 반응 최적온도은 각각 40, 45, $40^{\circ}C$이었고, 반은 최적 pH는 5.5, 6.0 그리고 6.0이었다. 효소의 열안정성은 30~$70^{\circ}C$에서 6시간 처리하였을 때 avicelase와 $\beta$-glucosidase는 $50^{\circ}C$ 이상에서 거의 실활하였고, CMCase는 $50^{\circ}C$에서 약 40%의 활성을 유지하였다. 효소의 안정성에 미치는 pH의 영향은 $25^{\circ}C$에서 24시간 처리하였을 때 avicelase와 CMCase는 PH 5.0~9.0 사이에서 안정하였으며, $\beta$-glucosidase는 pH 5.0~8.0 사이에서 안정하였다.

• 미생물학회지
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• 제14권2호
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• pp.84-94
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• 1976

Cellulase components, CMCase(Cx) and Avicelase$(C_1)$, were partially prueified, from the culture extract of a strain of Trichoderma koningii by column chromatography on DEAE-Sephadex A-50 and the step of gel filtration through Sphadex G-150, Optimum pH of CMCase was 5.2 and Avicelase showed the highest activity at pH 5.6 in acetate buffer. Optimal temperatures for activities of CMCase and Avicelase were $50^{\circ}C\;and\;60^{\circ}C, $ respectively. More than 70% of the activities of two enzymes were remained after heating at $60^{\circ}C$ for 30 min and Avicelase is more stable than any other fungal enzymes. The Michaelis constants, Km, of CMCase and Avicelase were 0.116% of CMC and 0.281% of avicel. And also the values of maximum velocity, Vmax, of CMCase and Avicelase were $23.20{\mu}g\;and\;2.54{\mu}g$ of reducing sugar per min. Of the metal ions tested against the activites of CMCase and Avicelase, $Cu^{++}, \; Hg^{++}, \;and\;Pb^{{++}$ are remarkably effective inhibitors. The molecular weights of Cx and $C_1$ component were estimated to be about 47, 000 and 61, 000 by gel filtration method.

• 미생물학회지
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• 제20권3호
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• pp.125-133
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• 1982

Mutational experiments were performed to imporve the cellulase productivity of Aspergillus phoenicis KU175, isolated from the southern part of Korea, as a high cellulase producer. By treatment ultra-violet light nad 4-NQO(4-Nitroquinoline-N-Oxide), mutation waas induced, and treatment ultra-violet light and 4-NQO (4-Nitroquinoline-N-Oxide), mutation was induced, and A.phoenicis KU175-115 was finally selected for its highest avicelase production. Avicelase production of the mutant was increased about 2 times compared with those of the wild strain. However, activities of other hydrolytic enzymes, such as amylase, protease and nuclease, of the mutant strain didn't show a marked difference compared with those of the nuclease, of the mutant strain didn't show a marked difference compared with the wild strain, except slight increase in ribonuclease activity and slight decrease in glucoamylase activity. Avicelases from the mutant strain selected were purified from wheat bran culture by successive salting out, followed by dialysis and column chromatography, and their charcteristics were compared with thosw of the wild strain. Avicelase was separated into three peaks in the mutant strain as well as in the case of wild strain. Avicelase II activity of the mutant strain was prominently higher than that of the wild strain, while avicelase I and III activities of those were equivalent. The optimal pH ranges and stability of avicelase II from the mutant strain were pH4-5 and pH3.5-6.0, respectively, as well as in the case of the wild strain. The optimal temperature and thermal stability of avicelase II from the mutant strain were $40{\sim}50^{\circ}C\;and\;20{\sim}55^{\circ}C$, respectively. These results were same as those of the wild strain. By the using of Eadie-Hofastee plot, $K_m\;and\;V_{max}$ of avicelase II from the mutant and the wild strain were calculated to be 2.29mg/ml and $4.84{\mu}g$ reducing sugar as glucose per min equally, from the line fitted to the data by the least square method. Activity of avicelase II from the mutant strain was slightly activated by $Mg^{++}\;but\;inhibited\;by\;Cu^{++}, \;Mn^{++}\;and\;Zn^{++}$, as well as in the case of the wild strain. Therefore, it was concluded that the mutant didn't induce the formation of another avicelase isozyme, or the changes in the properties of avicelase, but induce the changes in the productively of the same avicelase II by the action of regulatory gane.

• 한국균학회지
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• 제27권2호통권89호
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• pp.94-99
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• 1999

섬유소자원을 이용하기 위하여 S. rugosoannulata를 합성배지에서 배양하여 cellulase 생산 최적 배양조건을 검토한 결과는 다음과 같다. S. rugosoannulata에 의한 cellulase 생산은 세 효소 모두 $40^{\circ}C$가 최적온도이었고, Avicelase와 ${\beta}-glucosidase$는 pH 5.0, CMCase는 pH 4.0이 최적조건 이었다. 탄소원은 CMCase와 ${\beta}-glucosidase$는 xylose를 Avicelase는 maltose를 탄소원으로 했을 때 최고의 생산을 나타냈으며, xylose의 최적농도는 CMCase는 1.0%, Avicelase는 0.8%, ${\beta}-glucosidase$는 1.1%이었다. 질소원은 무기질소원으로 $NH_4Cl$ 첨가시 최고의 생산성을 나타냈으며, 최적농도는 CMCase는 0.3을 Avicelase와 ${\beta}-glucosidase$는 0.4%이었으며, 유기질소원으로는 malt extarct 첨가시 높은 생산성을 나타냈으며, 최적농도는 CMCase와 Avicelase는 1.0%, ${\beta}-glucosidase$는 1.3%이었다. 무기염류로는 $CoCl_2$ 첨가시 최고의 생산성을 나타냈으며, 최적농도는 세 효소 모두 0.35%이었다.

• 한국균학회지
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• 제12권4호
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• pp.133-140
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• 1984

Pleurotus sajor-caju JAFM 1017을 합성배지(合成培地)에 배양(培養)하여 배양중(培養中)에 생성(生成)된 섬유소(纖維素) 분해효소(分解酵素)의 성질(性質)을 검토(檢討)한 결과(結果), 작용최적(作用最適) pH는 avicelase가 pH5.5, CMCase는 pH4.5, ${\beta}-glucosidase$는 pH6.0이었고, pH안정(安定)범위는 avicelase는 $pH5.0{\sim}6.0$, CMCase는 $pH4.0{\sim}6.0$,${\beta}-glucosidase$는 $pH5.5{\sim}6.5$이었다. 최적온도(最適溫度)는 avicelase, CMCase ${\beta}-glucosidase$ 모두 $40^{\circ}C$이었고 열안정성(熱安定性)은 최적온도(最適溫度) 이하(以下)에서 안정성(安定性)을 보였으나 $50^{\circ}C$ 이상(以上)에서는 불안정(不安定)하여 avicelase는 $70^{\circ}C$, 10분(分)에 8.3%정도(程度)의 잔존활성(殘存活性)을 보였다. 효소(酵素)의 활성(活性)은 기질농도(基質濃度)가 증가(增加)함에 따라 증가(增加)하여 avicelase는 1%, CMCase는 0.7%, ${\beta}-glucosidase$ 0.1%까지 비례적(比例的)으로 증가(增加)하였으며 이들의 Km치(値)는 avicelase가 $30.77mg{\cdot}\;avicel/ml$, CMCase는 14.64mg CMC/ml, ${\beta}-glucosidase$는 5.1 3mg salicin/ml이었다. 반응시간(反應時間)에 따른 환원당(還元糖)의 생성(生成)은 Avicelase는 120분(分) CMCase와 ${\beta}-glucosidase$는 60분(分)까지 비례적(比例的)으로 증가(增加)하였다. 금속(金屬) ion의 영향(影響)은 $Ca^{2+}$은 $10^{-2}M$농도(濃度)에서 효소(酵素)의 활성(活性)을 증가(增加)시켰으나 $Hg^{2+},Ag^+$은 크게 저해(沮害)하였다.

• 미생물학회지
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• 제14권1호
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• pp.17-24
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• 1976

Celluloytic microorgnasims were isolated form the various sources and four of them were identified as Trichoderma roningi, Aspergillus niger, Penicillium chrysogenum, and Streptomyces sp. The induction of extracellular5 cellulase of these species in the liquid culture media containing carboxymethylcellulose (CMC) or Avicel as inducer showed that CMC was a better effective inducer for the production of CMCase(Cx cellulase component) as well as Avicelase(C$_{1}$ cellulase component) than Avicel. It is believed that certain hydrolysis products of cellulose(CMC) could serve as an inducer for an enzyme synthesis. In T. roningi, Asp. niger, and Strptomyces sp., the optimum temperature of CNCase on CMC-culture medium was 50.deg. but temperature around 40.deg.C was found to be optimum for the activities of CMCase prepared from P.ehrysogenum. The optimum temperature for Avicelase activitiles on Avicel-culture media of T. roningi and P. chrysogenum was $40{\circ}C$ whereas temperature $50{\circ}C$ was found to be optimum for Avicelase from A.niger and Streptomyces sp. The optimal activities of these CNCase and Avicelase prepared from. T. ronigi, Pen.chrysogenum and Streptomyces sp. were found similarly to be at pH's around 5.4 and 6.0 while pH 4.8 was optimum for the activities of CMCase and Avicelase from A.niger, indicating that A.niger in acidic media would yield an enzyme of high activity.

• 한국균학회지
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• 제15권1호
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• pp.29-37
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• 1987

창덕궁의 유물고에 소장되어 있는 섬유질 문화재로부터 섬유소 분해균을 분리하기 위하여 두 단계의 선별을 거쳐서 섬유소 분해능이 강력한 Cellulolytic fungus를 분리하였다. 이 균주는 형태학적인 분류기준을 통하여 Aspergillus clavatus로 동정되었다. 이 균주에서 CMC 액체배지에서의 CMCase, avicelase와 salicinase의 생성은 $30^{\circ}C$의 shaking culture 조건에서 5 일에 가장 높았으며, 그 이후는 점차 감소하였다. crude extracellularenzyme 을 $70%(NH_4)_2SO_4$ 포화용액에서 침전시켜 20mM acetate buffer(pH 6.0)로 dialysis시킨후 효소에 대한 여러가지 성질을 비교하였다. CMCase와 avicelase의 최적 pH는 모두 pH6.0으로 중성에 가까왔으며 두 효소의 최적온도는 모두 $50^{\circ}C$이었다. 효소의 열에 대한 안정도에서는 CMCase, avicelase 모두 $30-50^{\circ}C$에서 안정성을 유지하였고 또한 효소의 활성에 미치는 기질농도의 영향을 보먼 CMCase는 1.5%에서 avicelase는 2.2%에서 최고의 활성을 나타냈다. 금속이온의 영향을 살펴본 결과 $Mg^{++}$와 $Ca^{++}$는 5mM에서 효소의 활성이 최고에 달하였으며 $Cu^{++}$와 $Zn^{++}$에서는 농도가 증가할수록 효소의 활성이 저해되었다. 또한 이 균주의 섬유소 분해효소는 2-mercaptoethanol, $I_2$, $NaN_3$ 등의 저해제에 의하여 활성이 크게 저해되었다.

• KSBB Journal
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• 제17권4호
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• pp.381-387
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• 2002

본 연구의 목적은 기존의 단일 균주보다 두 종의 서로 다른 균주를 복합시켰을 때 두 균주간의 상호 보완적인 면을 이용하여 효소생산을 극대화하기 위함이며, 다음과 같은 결론을 얻었다. 실험에서 사용된 T viride와 FB01는 최적 PH와 온도가 비슷하여 혼합배양이 가능하였으며, 이들의 복합균주를 개발하는데 성공하였다. 즉 두 균주는 회분배양 하였을 때, 서로의성장속도에 차이가 있었기 때문에 두 균주의 생장균형을 유지하기 위해 접종시기를 조절할 필요가 있었다. 접종시기를 변화시킴으로서 두 균주간의 상호작용으로 인해 단일 균주일때보다 CMCase, $\beta$-glucosidase 및 avicelase의 활성이 우수했으며 또한 계대배양을 통한 두 균주의 활성유지에 관한 실험에서는 pH 조절을 통해 $\beta$-glucosidase 활성이 최고 3.2배까지증가함을 알 수 있었다. 따라서 배양액내에서 초기 pH의 영향은 생산된 cellulase complex의 다양한 구성요소의 상대적인 농도를 결정하는 실질적인 요인으로 판명된다. pH 4.5에서의 enzyme activity는 12.5 U/mL CMCase, 2.46 U/mL If-glucosidase, 0.93 U/mL avicelase였으며 개발당시의 복합균주활성인, 2.04 U/ml CMCase, 0.78U/ml $\beta$-glucosidase, 0.2 U/mL avicelase과 비교할 때 각각 6, 3.2, 4.7배의 높은 활성을 보였다. 복합균주의 탄소원으로 볏짚을 이용했을 때 복합균주에 의한 효소생산이 단일 균주인 T. virile나 FB01보다 훨씬 높게 나타났으며, 이때의 볏짚의 최적 농도는 5%(w/v)였다. 이러한 배양조건에서 복합균주를 장기적으로 계대배양하면서 효소의 생산성 향상과 복합균주의 안정성을 관찰할 필요가 있다고 사료된다.

• 미생물학회지
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• 제15권3호
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• pp.113-121
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• 1977

The 685 strains of Korean Sspergilli are isolated, cultured purely, and their Avicelase, CMCase and Salicinase activities are measured, in order to select the best strains exhibiting predominant cellulase activities, and to survey their cellulase activities in taxonomical and ecological viewpoints. Strains No.175, 255 and 254 are selected as the best Avicelase producing strains, strains No.131, 151 and 116 are selected as the best CMCase producing strains, and strains nO.456, 457 and 253 are selected as the best Salicinase producing strains. A niger group and A, clavatus group exhibited the highest activities of Avcelase and Salicinase, respectively. A.iniger group and A, clavatus groups are showed the highest activites of CMCase. Among the different species tested, the activities of Avicelase, CMCase andSalicinase are highest in A.phoenicis, A.clavatus, and A. japonicus and A.giganteus, respectively. Cellulase activities of Aspergilli from the inland regions of Korea are higher, more or less, than those of the strains from the other regions. Avicelase and CMCase activities of Aspergilli isolated from bread and Korean cake are relatively higher, and Salicinase activities of the strains isolated from the cereals are higher than those of the strains from the other habitat substrate.