• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.514-523
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• 2017

Carbon catabolite repression is a crucial regulation mechanism in microorganisms, but its characteristic in Rhizopus is still unclear. We extracted a carbon regulation gene, cre, that encoded a carbon catabolite repressor protein (CRE) from Rhizopus stolonifer TP-02, and studied the regulation of CRE by real-time qPCR. CRE responded to glucose in a certain range, where it could significantly regulate part of the cellulase genes (eg, bg, and cbh2) without cbh1. In the comparison of the response of cre and four cellulase genes to carboxymethylcellulose sodium and a simple carbon source (lactose), the effect of CRE was only related to the concentration of reducing sugars. By regulating the reducing sugars to range from 0.4% to 0.6%, a glucose-containing medium with lactose as the inducer could effectively induce cellulases without the repression of CRE. This regulation method could potentially reduce the cost of enzymes produced in industries and provide a possible solution to achieve the largescale synthesis of cellulases.

• Asian-Australasian Journal of Animal Sciences
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• 제12권4호
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• pp.531-536
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• 1999

This study was conducted to compare the effects of lactic acid bacteria (LAB) or LAB+cellulases on the cell wall compositions and the in vitro dry matter digestibility (IVDMD) of Rhodesgrass (RG) and Italian ryegrass (IRG) silages. LAB (Lactobacillus cassei) at a concentration of $10{\times}10^5\;cfu.g^{-1}$ fresh forage was added to all ensiling samples (except the untreated control) of RG and IRG. The cellulases used were Acremoniumcellulase (A), Meicelase (M) or a mixture of both (AM). Each cellulase was applied at levels of 0.005, 0.01 and 0.02 % fresh sample. The samples were incubated at 20, 30 and $40^{\circ}C$ for about 2 months of storage. LAB inoculation did not affect cell wall components or IVDMD of both the RG and IRG silages, but LAB+cellulase treatments did. Increasing the amount of cellulase addition resulted in further decreases of cell wall concentrations. This reduction more markedly occurred with cellulases A and AM than it did with cellulase M. Cell wall components losses were higher in the IRG silages than in the RG silages. LAB+cellulase treatments decreased IVDMD of the RG silages, but had no effect on the IRG silages. The different effect of LAB+cellulase treatments on cell wall degradation and IVDMD of the RG and IRG silages suggested that RG contains more structural carbohydrates, which were difficult to degrade with cellulase, than did IRG.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2005년도 2005 Annual Meeting & International Symposium
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• pp.245-247
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• 2005

A cellulolytic fungus isolated from Agave plantation in northeastern Thailand was identified as Acrophialophora nainiana. The fungus was capable of growing at pH between 3 - 7 and 25 - 45 $^{\circ}C$, with the optimum conditions at pH 5.0 and 40 $^{\circ}C$. The wild isolate produced cellulases, comprising of exoglucanase (0.019 U/mg protein), endoglucanase (0.366 U/mg protein), and ${\beta}$-glucosidase (0.001 U/mg protein). Mutations with UV and NTG produced the UV 10-2 mutant with cellulases activities including exoglucanase (0.093 U/mg protein), endoglucanase (0.585 U/mg protein), and ${\beta}$-glucosidase (0.013 U/mg protein). Purification of the enzymes with ultrafiltration, ammonium sulfate precipitation, and ion-exchange chromatography yielded the maximal cellulase specific activities of 2.736 U/mg protein (exoglucanase), 0.235 U/mg protein (endoglucanase), and 0.008 U/mg protein (${\beta}$-glucosidase). The mutant's cellulases were the most active at pH 5.0 and 60 $^{\circ}C$. Ion-exchange chromatography revealed that A. nainiana UV 10-2 cellulases were comprised of two peaks with one peak showing the single endoglucanase activity while the other peak showed a mixture of the three enzyme activities. Production of A. nainiana UV 10-2 cellulases using banana leaf stalk as the sole carbon source gave comparable yields to that of the pure ${\alpha}$-cellulose. The enzymes were used in the simultaneous saccharification and fermentation (SSF) of plant residue (Coix aquatica) along with Kluveromyces marxianus to produce ethanol. Moreover, when the enzymes were used in the bioscouring process of fabric, the desiravle traits of textile processing including immediate water absorbency, increased in whiteness and reduction of yellowness of the treated fabric were observed.

• 한국버섯학회지
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• 제14권4호
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• pp.127-135
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• 2016

Cellulose is the most abundant organic polymer constituent of the cell wall of green plants and of various forms of algae. The complexity of lignocellulosic biomass is a major challenge in industrial research. Most mushroom species that naturally grow on soil or wood possess cellulases and the corresponding enzymatic system and, potential candidates for the direct bioconversion of softwood polysaccharides into fermentable sugars. However, there have been fewer studies on mushroom cellulases than on fungi such as Trichoderma spp., exploit the full potential of mushroom cellulases. This review will focus on the current status ofmushroom cellulase research and applications and will provide insight into promising future prospects.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.122-129
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• 2017

Termites are destructive to agriculture, forestry, and buildings, but they can also promote agro-ecosystem balance through the degradation of lignocellulose. Termite-triggered cellulose digestion may be clarified through microbial metabolism of cellulose products. In the present study, we characterized the activities of cellulase and its three components synthesized by the cellulase-producing fungal strain HDZK-BYTF620 isolated from the gut of Odontotermes formosanus. The protein components of cellulases were synthesized by strain HDZK-BYTF620, which were isolated and characterized using polyacrylamide gel electrophoresis, and the expression of the cellulases was studied at the proteome level.

• Asian-Australasian Journal of Animal Sciences
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• 제12권4호
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• pp.525-530
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• 1999

Prior to ensiling Rhodesgrass (RG) and Italian ryegrass (lRG) were treated with lactic acid bacteria (LAB) or with LAB+cellulases to compare their fermentation characteristics and chemical compositions. LAB (Lactobacillus casei) was added to all ensiling materials (except the untreated control) of RG and IRG at a concentration of $1.0{\times}10^5\;cfu.g^{-1}$ fresh forage. The enzymes used were Acremoniumcellulase (A), Meicelase (M) or a mixture of both (AM). Each enzyme was applied at levels of 0.005, 0.01 and 0.02 % of fresh forage. The silages with each treatment were incubated at 20, 30 and $40^{\circ}C$ and stored for about 2 months. While no marked differences were found between the RG and IRG silages with various treatments on dry matter (DM), volatile basic nitrogen (VBN) and water soluble carbohydrate (WSC) contents, there were significant differences in pH value, and lactic acid and butyric acid contents. LAB inoculation did not affect the fermentation characteristics of either the RG or IRG silages. The combined treatments of LAB+cellulases improved the fermentation quality of both the RG and IRG silages as evidenced by the decrease in pH value and increase in lactic acid content. Increasing the amount of added cellulase resulted in a decrease in pH value and an increase in lactic acid content in both the RG and IRG silages. Cellulases A and AM had a greater effect than cellulase M on the fermentation quality of the RG and IRG silages. Incubation temperatures of 30 and $40^{\circ}C$ appeared to be more appropriate environments for stimulating good fermentation than $20^{\circ}C$.

• 미생물학회지
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• 제13권3호
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• pp.85-90
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• 1975

Crude cellulases freshly prepared from cultures of Aspergillus niger, Prnicillum motatum, Trichoderma vride 16273 and Trichoderma viride 16374 were assayed on 4 different substrates including Na-CMC, cellulose powder, starch and sucrose. Enzyme prepared from A. niger contained highly active hydrolytic enzymes of the 4 substrates assayed. P. notatum [yielded relatively lower amount of cellulase but the extracts were also highly reactive on starch and sucrose. Trichoderma viride 16274 yielded very little cellulase and invertase, but the extracts showed a high degree of amylase activity. Trichoderma viride 16374, however, yielded collulase comparable to that of Penicillium notatum, but lower activities of amylase and invertase were seen. Commercial cellulases prepared from Penicillium notatum (cellulase[K]) and Trichoderma viride(cellulase[J]) indicated enzyme activities closely parallel to the crude enzymes freshly prepared from fungus cultures. The optimum pH's of cellulolytic activities of cellulase[K] and cellulase[J] were 4.0 and 5.0 respectively. The optimum temperatures of the cellulolytic activities of cellulase[K] and cellualse[J] were 4.0 and 5.0 respectively. The optimum temperatures of the cellulolytic activities of cellulase [K] and cellulase [J] were $60{\circ}C$ and $50{\circ}C$ respectively. Assuming the average molecular weight of Na-CMC is about 115,000, the Km values of cellulase [K] and cellulase[J] were found to be $3.3{\times}10^{-5}/nM$ and $3.3{\times}10^{-4}/nM$ respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제31권12호
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• pp.1913-1922
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• 2018

Objective: To find out ways of improving fermentation quality of silage, the comparative analysis of fermentation characteristics and in vitro digestibility of tropical grasses silage applied with cellulases produced from Acremonium or Tricoderma species were studied in Thailand. Methods: Fresh and wilted Guinea grass and Napier grass silages were prepared with cellulases from Acremonium (AC) or Trichoderma (TC) at 0.0025%, 0.005%, and 0.01% on a fresh matter (FM), and their fermentation quality, chemical composition and in vitro digestibility were analyzed. Results: All silages of fresh Napier grass were good quality with lower pH, butyric acid, and ammonia nitrogen, but higher lactic acid content than wilted Napier grass and Guinea grass silage. Silages treated with AC 0.01% had the best result in terms of fermentation quality. They also had higher in vitro dry matter digestibility and in vitro organic matter digestibility at 6 and 48 h after incubation than other silages. Silages treated with lower levels at 0.005% or 0.0025% of AC and all levels of TC did not improve silage fermentation. Conclusion: The AC could improve silage fermentation and in vitro degradation of Guinea grass and Napier grass silages, and the suitable addition ration is 0.01% (73.5 U) of FM for tropical silage preparation.

• 한국식품과학회지
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• 제30권2호
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• pp.245-252
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• 1998

키토산 올리고당 생산에 이용 가능한 경제성 있는 키토산 분해 효소를 탐색하였다. Cellulase, ${\beta}-1,4-glucosidase,\;protease$ 및 상업용 효소류를 대상으로 조제 키토산에 대한 분해력을 시험한 결과 Trichoderma viride, T. reesei 유래의 cellulase 및 상업용 효소인 Celluclast (T. reesei 유래 cellulase)가 우수한 키토산 분해능력을 나타내었다. 키토산에 대한 반응성은 T, reesei 유래 cellulase가 T. viride 유래 cellulase 보다 강하였으며 이들의 적정 키토산 분해조건은 pH 5.0이었고 적정 온도는 T. viride 유래 celluclast는 $45^{\circ}C$, T. reesei 유래 cellulase 및 Celluclast는 $55^{\circ}C$로 나타났다. 또한 dose reponse시험에서 이들 효소 모두 enz./chitosan비=0.1이 적절하였으며 T. reesei 유래 효소 및 Celluclast는 키토산 농도가 3%일 때 최대의 활성을 나타내었다. 한편, 상업용 효소인 Celluclast를 이용한 키토산 올리고당의 생산 가능성을 시험하기 위하여 3% 키토산 용액(310cp)에 Celluclast를 1% 첨가하고 $pH\;5.0,\;55^{\circ}C$에서 시간별로 반응시켰을 때, 점도는 초기30분경에 5.51cp로 98% 이상 감소하였으며 50% ethanol 가용물의 수율은 분해 15시간에 70%로 최대를 보였다. 총환원당의 함량은 분해시간이 경과함에 따라 증가하였고 반응 2시간부터 13.5% 내외의 수준을 유지하였다. 올리고당의 조성은 15시간 분해물에서 균일한 분포와 높은 함량을 보였으며 항종 양활성을 나타내는 것으로 알려진 6량체 키토산 올리고당의 생성량은 기존의 산 분해방법의 약 4배에 해당되는 8.0%로 나타났다. 따라서 T. viride 및 T. ressei 유래의 cellulase는 99% 이상의 탈아세틸화도를 가지는 키토산에 대하여 분해활성을 나타내고 특히 T. reesei유래의 상업용효소는 지금까지 효소가격이 너무 높아 활용되지 못한 chitosanase를 대신하여 키토산 올리고당을 경제성 높게 생산하는 데 이용될 수 있을 것으로 기대되었다.

• Animal Bioscience
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• 제34권5호
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• pp.867-879
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• 2021

Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.