• Journal of Microbiology and Biotechnology
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• 제2권1호
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• pp.50-55
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• 1992

A genomic library of a mesophilic cellulolytic anaerobe, Clostridium sp. KCTC 8440 DNA was constructed in Escherichia coli using plasmid pUC9. Clones of E. coli exhibiting carboxymethyl cellulose-hydrolyzing activity (CMCase) were isolated and divided into seven types based on the restriction enzyme patterns of recombinant plasmids. E. coli strains carrying type A genes showed activity on carboxymethyl cellulose about 7-8 times greater than clones carrying genes of other types. Restriction maps of the cloned DNA fragments were determined, and homologies between them were investigated. The results suggest that Clostridium sp. KCTC 8440 has seven distinct CMCase genes.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1196-1206
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• 2014

A metagenomic fosmid library was constructed using genomic DNA isolated from the gut microflora of Hermetia illucens, a black soldier fly. A cellulase-positive clone, with the CS10 gene, was identified by extensive Congo-red overlay screenings for cellulase activity from the fosmid library of 92,000 clones. The CS10 gene was composed of a 996 bp DNA sequence encoding the mature protein of 331 amino acids. The deduced amino acids of CS10 showed 72% sequence identity with the glycosyl hydrolase family 5 gene of Dysgonomonas mossii, displaying no significant sequence homology to already known cellulases. The purified CS10 protein presented a single band of cellulase activity with a molecular mass of approximately 40 kDa on the SDS-PAGE gel and zymogram. The purified CS10 protein exhibited optimal activity at $50^{\circ}C$ and pH 7.0, and the thermostability and pH stability of CS10 were preserved at the ranges of $20{\sim}50^{\circ}C$ and pH 4.0~10.0. CS10 exhibited little loss of cellulase activity against various chemical reagents such as 10% polar organic solvents, 1% non-ionic detergents, and 0.5 M denaturing agents. Moreover, the substrate specificity and the product patterns by thin-layer chromatography suggested that CS10 is an endo-${\beta}$-1,4-glucanase. From these biochemical properties of CS10, it is expected that the enzyme has the potential for application in industrial processes.

• 한국미생물·생명공학회지
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• 제31권2호
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• pp.124-128
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• 2003

대전광역시 근교의 야산과 들판 등지에서 썩은 나뭇잎, 짚, 흙을 채취하여 각각을 배양한 다음 Congo red test에 의해 cellulase 활성을 보이는 균주를 선별하였다. Genomic DNA를 분리한 후 PCR을 수행하여 DNA sequence를 Gene Bank를 통해 분석한 결과 A. tubingensis로 밝혀졌다. 이것을 배양하여 상등액을 crude enzyme으로 사용하여 온도와 pH를 달리하면서 효소의 활성정도를 측정하였다. 대조균주로 A. oryzae KCTC 6291를 이용하였고, 본 연구를 통하여 분리한 균주인 A. tubingensis가 생산하는 cellulase는 A. oryzae의 cellulase에 비하여 각각 다른 온도와 pH에서 높은 안정성을 보여주었다. A. tubingensis는 각각의 온도에서 활성의 정도가 비슷했으며, 45$^{\circ}C$, 55$^{\circ}C$에서 높은 활성을 나타내고 있지만, 고르게 활성이 나타났다. 또한 pH 12.0에서 가장 높은 활성을 보여 주었고, pH 2.0, 3.0, 4.0에서는 양쪽 모두 거의 활성이 없었으며, 중성, 염기성에 대해서 활성에는 큰 변화가 없었다. 따라서, 분리 동정한 A. tubingensis는 온도와 pH에서 고르게 활성을 나타내므로 생균제로 활용할 수 있는 범위가 클 것으로 여겨진다.

• 한국펄프종이공학회:학술대회논문집
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• 한국펄프종이공학회 1999년도 Pre-symposium of the 10th ISWPC Recent Advances in Paper Science and Technology
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• pp.291-295
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• 1999

Coprinus cinereus 2249 that is a kind of basidiomycetes constitutively produced alkaline carboxymethyl cellulase (CMCase), filter paper cellulase (FPase) and xylanase. Crude enzymes prepared with optimal conditions showed higher FPase activity than CMCase activity. The FPase was most active at pH 9 at 50$^{\circ}C$. When applied on deinking of the old newsprint (ONP), it increases the freeness and brightness due to effect of hydrolysis at 0.1% enzyme concentration. Also, The physical properties of deinked pulp were improved.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.362-368
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• 2001

The effects of water soluble materials in paper mill sludge on cellulase and $\beta$-glucosidase activities were studied while the optimization of enzyme system for hydrolysis of the paper mill sludge for production of glucose was made. Water soluble materials in the paper mill sludge showed stimulatory effect on carboxymethyl cellulose (CMC) activity, inhibitory effect on filter paper (FP) activity, and no effect on avicelase and $\beta$-glucosidase activities. CMC and ${\beta}$-glucosidase activities at 5 and 10, 5 or 10 and 10, and 10 and 10 U/ml were optimal for hydrolysis of 5, 10, and 20% of the paper mill sludge, respectively.

• 미생물학회지
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• 제22권2호
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• pp.97-101
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• 1984

To investigate the biodegradation pattern of rice straw, mainly composed of cellulose, hemicellulose and lignin components, by the isolate stran Bacillus subtilis $DO_4$, the change of cell population was observed on CMC (carboxymethyl cellulose), larch wood xylan and lignosulfonate as a carbon source respectively. Also, the transition pattern of enzyme activities of cellulase and xylanase and lignin contents was measured on rice straw and mixed substrate according to growth. The results in these experiments revealed that xylanase activity was first appeared and cellulase activity in the next, while lignin component was almost not changed through the culture period.

• 한국미생물·생명공학회지
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• 제50권1호
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• pp.122-125
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• 2022

The cellulose-fed microbial fuel cell (MFC) is a biotechnological process that directly converts lignocellulosic materials to electricity without combustion. In this study, the cellulose-fed, MFC-integrated thermophilic bacterium, Bacillus sp. WK21, with endoglucanase and exoglucanase activities of 1.25 ± 0.08 U/ml and 0.95 ± 0.02 U/ml, respectively, was used to generate electricity at high temperatures. Maximal current densities of 485, 420, and 472 mA/m² were achieved when carboxymethyl cellulose, avicel cellulose, and cellulose powder, respectively, were used as substrates. Their respective maximal power was 94.09, 70.56, and 89.30 mW/m³. This study demonstrates the value of the novel use of a cellulase-producing thermophilic bacterium as a biocatalyst for electricity generation in a cellulose-fed MFC.

• 한국균학회지
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• 제42권1호
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• pp.40-49
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• 2014

우리나라 산림에서 수집한 목재부후성 버섯 균주들에 대한 배지 및 온도별 균사생장 특성을 고체평판배지 상에서 조사하였다. 대부분의 균주들은 malt extract agar (MEA)나 sabouraud dextrose agar (SDA)보다는 PDA (potato dextrose agar) 배지에서 균사 생장력이 더 높았다. 몇몇 균주들을 제외하고, 균주들의 대부분은 PDA 배지 상에서 배양 시 $25^{\circ}C$에서 가장 잘 생장하였으며, 고온($30^{\circ}C$)보다는 저온($10^{\circ}C$)에서 균사 생장이 저조하였다. 2종류의 서로 다른 세포외효소 활성에 각각 특이적으로 반응할 수 있는 기질이 함유된 고체평판배지 상에서 모든 시험균은 carboxymethyl cellulase 활성과 laccase 활성을 나타냈다.

• 한국균학회지
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• 제25권1호통권80호
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• pp.68-76
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• 1997

섬유소자원의 이용을 목적으로 담자균류인 T. trogii를 실험 균주로 하여 합성배지에서 cellulase 생성 최적 배양조건을 검토해 본 결과는 다음과 같다. 합성배지에서 T. trogii에 의한 cellulase 생산은 $30{\sim}35^{\circ}C,\;pH\;4.0{\sim}6.0,\;and\;11{\sim}15$일 배양시 가장 높았다. 탄소원은 Avicelase와 ${\beta}-glucosidase$의 경우 CMC를, CMCase는 cellulose를 탄소원으로 했을 때 최고의 생산성을 보였으며, CMC의 최적 농도는 세 효소 모두 3%이었다. 질소원은 ammonium tartrate 첨가시 cellulase 생산이 높았으며, 그 최적 농도로 Avicelase와 ${\beta}-glucosidase$는 0.03N, CMCase는 0.04 N이었다.

• 생명과학회지
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• 제22권4호
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• pp.437-446
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• 2012

한우의 반추위에서 게놈 DNA를 분리하여 메타게놈 은행을 구축한 다음 섬유소분해효소를 암호화하는 유전자를 클로닝 및 유전자를 선별하였다. 선별된 유전자의 DNA 염기서열 및 아미노산 서열을 분석하고 유전산물의 생화학적인 특성을 조사하였다. $cel$5C 유전자는 1,125 bp로 374개의 아미노산 잔기를 가진 단백질을 암호화하였으며 이 단백질 분자량은 42 kDa이었다. 이 효소의 최적 pH는 4 근방이었으며 최적 온도는 $50^{\circ}C$ 부근이었다. $cel$5C 유전자의 internal primer를 사용하여 인공적으로 배양할 수 있는 49종의 반추세균에서 분리한 게놈 DNA을 주형으로 PCR 분석한 결과 해당하는 밴드를 확인할 수 없었다. Cel5C는 현재로서는 배양할 수 없는 반추 미생물로 추정된다.