• 한국균학회지
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• 제31권2호
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• pp.77-83
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• 2003

천연자원인 cellulose를 이용하기 위하여 섬유소분해효소의 생산성이 강한 Roseofomes subflexibilis의 배양학적 특성을 조사한 결과는 다음과 같다. 1.섬유소분해효소의 생산 및 균사체 생장에 적합한 탄소원은 glucose인 것으로 나타났으며, 균사체배양의 탄소원은 saccharose로 최적농도는 1.2%였다. 2. 균사체배양의 유기질소원은 yeast extract이었으며. 최적농도는 1.5%이었다. 또한 무기질소원은 $NH_{4}H_{2}PO_{4}$를 잘 이용하는 것으로 나타났고, 최적농도는 1.1% 첨가시 균사체의 생장이 좋았다. 3. 균사체배양의 아미노산의 영향을 조사한 결과 aspartic acid가 가장 좋았으며, 최적농도는 0.1%이었다. cysteine 첨가시 효소생산과 균사체의 생장 모두에서 효과적이었다. 4.무기염류의 영향은 $Al_{2}(SO_{4})_{3}$ 첨가시 효과적이었으며 최적농도는 0.1 M이었다. 5.배양온도는 $30{\circ}C$였으며, 최적 pH는 4.0인 것으로 나타났다.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.383-389
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• 1990

Penicillium verruculosum 섬유소 분해효소의 유도물질인 KC-flock 및 cellobiose octaacetate(COA)를 유일한 탄소원으로 포함하는 배지에서 21일간 배양하면서 배양여액 중의 섬유소 분해효소 각 성분의 활성도와 전기영동상의 단백질 양상을 검토하였다. 배양여액의 중의 총단백질 생성 및 섬유소 분해활성의 유도 효과는 COA 배지가 다른 두 배지의 3배 이상이었다. 배양기간의 증가에 따라 배양여액 중의 총단백질량이 증가되었는데 CMC 분해활성이나 $\beta$-glucosidase 활성의 증가보다는 avicel 분해활성도의 증가가 총단백질량의 증가와 유사하다.

• 미생물학회지
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• 제23권1호
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• pp.25-33
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• 1985

Cellulomonas flavigena KIST 321이 생산하는 균체외 cellulose 분해효소를 gel filtration법과 ion-exchange chromatography법으로 3종의 다른 효소를 분리하여 이들을 A, B, 및 C 효소로 명명하였다. 분리된 각 효소를 결정적 기질에 처리하여 일어나는 기질의 구조변화를 적회전 분광법과 X-ray crystallography법으로 측정하여 다음 결론을 얻었다. B효소는 결정성 cellulosem이 구성단위인 glucopyranose의 불안정화로 그 결정도를 감소시키는 $C_1$형의 효소이며 A 및 C 효소는 $C_x$형의 효소로 B 효소의 반응생성물에 작용하여 glucose를 생산하였다. 이들 각 효소의 작용에서 본균의 cellulase의 작용기작을 고찰하였다.

• Korean Chemical Engineering Research
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• 제52권3호
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• pp.302-306
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• 2014

목질계 분해효소 활성 증대를 위해 밀짚을 이용한 고체발효에서 주요 발효인자의 최적화를 수행하였다. Trichoderma reesei와 Aspergillus niger를 이용한 혼합배양에서 고체발효에 주요한 영향을 미친다고 알려진 배양온도, pH, 수분함량과 고체기질 크기를 순차적 최적화를 진행하였다. 실험에 적용 된 인자 모두 목질계 분해효소 활성에 유의한 효과를 주었으며, 발효온도 $40^{\circ}C$, pH 7, 수분함량 75%와 고체기질 크기 0.25~0.5 mm가 목질계 분해효소 생산을 위한 최적 조건임을 알 수 있었다. 최적조건 하에서 밀짚을 이용한 고체발효를 수행하였을 때, 효소활성 기준 cellulase 10.3 IU, endoglucanase 100.3 IU, ${\beta}$-glucosidase 22.9 IU와 xylanase 2261.7 IU/g dry material을 배양 96시간에 확인할 수 있었다. 본 결과는 기존 효소활성 대비 각각 72.6, 48.7, 55.2와 51.9% 증가한 수치로 혼합배양과 순차적 최적화를 적용하여 효과적인 목질계 분해효소 활성 증대가 가능함을 확인하였다.

• 한국균학회지
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• 제14권3호
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• pp.225-229
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• 1986

Lenzites betulina(Fr.)에서 효소 분획을 추출하여 carboxymethyl cellulose에 대한 분해 작용을 실험하였다. carboxymethyl cellulase의 활성도는 1.65 unit/ml였다.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1670-1679
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• 2020

The substantial use of fungal enzymes to degrade lignocellulosic plant biomass has widely been attributed to the extensive requirement of powerful enzyme-producing fungal strains. In this study, a two-step screening procedure for finding cellulolytic fungi, involving a miniaturized culture method with shake-flask fermentation, was proposed and demonstrated. We isolated 297 fungal strains from several cellulose-containing samples found in two different locations in Thailand. By using this screening strategy, we then selected 9 fungal strains based on their potential for cellulase production. Through sequence-based identification of these fungal isolates, 4 species in 4 genera were identified: Aspergillus terreus (3 strains: AG466, AG438 and AG499), Penicillium oxalicum (4 strains: AG452, AG496, AG498 and AG559), Talaromyces siamensis (1 strain: AG548) and Trichoderma afroharzianum (1 strain: AG500). After examining their lignocellulose degradation capacity, our data showed that P. oxalicum AG452 exhibited the highest glucose yield after saccharification of pretreated sugarcane trash, cassava pulp and coffee silverskin. In addition, Ta. siamensis AG548 produced the highest glucose yield after hydrolysis of pretreated sugarcane bagasse. Our study demonstrated that the proposed two-step screening strategy can be further applied for discovering potential cellulolytic fungi isolated from various environmental samples. Meanwhile, the fungal strains isolated in this study will prove useful in the bioconversion of agricultural lignocellulosic residues into valuable biotechnological products.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.482-493
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• 2005

Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.

• Asian-Australasian Journal of Animal Sciences
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• 제11권5호
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• pp.538-544
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• 1998

One hundred and twenty-eight finishing pigs (51.3 kg average initial body weight) were used to determine the effects of adding cellulase enzymes and lactobacillus acidophilus to sorghum-based diets on growth performance, carcass merit, and nutrient digestibility in finishing pigs. Treatments were: 1) corn-soybean meal-based positive control; 2) sorghum-soybean meal-based negative control; 3) Diet 2 with celluloytic enzymes; and 4) Diet 2 with a bacterial feed additive (lactobacillus acidophilus). There was a trend for greater average daily gain (ADG) in pigs fed com versus the sorghum treatments for day 0 to 28 (p < .09), but there was no effect of treatment (p > .15) on overall ADG (i.e., day 0 to 63). Feed consumption was not affected by treatment during the experiment (p > .19). Pigs fed the corn-soybean meal-based diet had 3.5% greater overall gain/feed than pigs fed the other diets (p < .009). Dressing percentage was not affected by treatment (p > .22), but there was a trend for backfat thickness at the last rib to be greater for pigs fed com versus the sorghum treatments (p < .09). Pigs fed the sorghum treatments had 1 % greater fat free lean index (p < .10) compared to pigs fed the corn-soybean meal-based positive control. Pigs fed com had greater apparent digestibilities of DM, N, and GE than pigs fed the sorghum treatments (p < .03), and greater DE intake (p < .07) suggesting that the increased carcass fatness for pigs fed the corn-based control diet resulted from greater energy status of those pigs. In conclusion, pigs fed the corn-soybean meal-based control diet had no improved growth performance but tended to be fatter than pigs fed sorghum. Adding cellulolytic enzymes or a bacterial feed additive to diets for finishing pigs did not affect growth performance, carcass merit, or nutrient utilization.

• Asian-Australasian Journal of Animal Sciences
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• 제27권7호
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• pp.946-950
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• 2014

Bacillus licheniformis was grown in minimal nutrient medium containing 1% (w/v) of distillers dried grain with soluble (DDGS), palm kernel meal (PKM), wheat bran (WB) or copra meal (CM), and the enzyme activity of endoglucanase, ${\beta}$-glucosidase, xylanase and reducing sugars was measured to investigate a possibility of using cost-effective agricultural residues in producing cellulolytic and hemicellulolytic enzymes. The CM gave the highest endoglucanase activity of 0.68 units/mL among added substrates at 48 h. CM yielded the highest titres of 0.58 units/ml of ${\beta}$-glucosidase, compared to 0.33, 0.23, and 0.16 units/mL by PKM, WB, and DDGS, respectively, at 72 h. Xylanase production was the highest (0.34 units/mL) when CM was added. The supernatant from fermentation of CM had the highest reducing sugars than other additional substrates at all intervals (0.10, 0.12, 0.10, and 0.11 mg/mL respectively). It is concluded that Bacillus licheniformis is capable of producing multiple cellulo- and hemicellololytic enzymes for bioethanol production using cost-effective agricultural residues, especially CM, as a sole nutrient source.

• Mycobiology
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• 제42권2호
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• pp.181-184
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• 2014

To the best of our knowledge, this is the first report on thermophilic fungi isolated in Korea. Three species of thermophiles were isolated from compost and were identified as Myriococcum thermophilum, Thermoascus aurantiacus, and Thermomyces lanuginosus. They can grow at temperatures above $50^{\circ}C$ and produce high levels of cellulolytic and xylanolytic enzymes at high temperatures. Notably, the considerable thermostability of the endo-glucanase produced by T. aurantiacus has made the fungus an attractive source of industrial enzymes.