• Asian-Australasian Journal of Animal Sciences
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• 제22권11호
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• pp.1555-1565
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• 2009

Anaerobic rumen fungi have been regarded as good genetic resources for enzyme production which might be useful for feed supplements, bio-energy production, bio-remediation and other industrial purposes. In this study, an expressed sequence tag (EST) library of the rumen anaerobic fungus Neocallimastix frontalis was constructed and functional genes from the EST library were analyzed to elucidate carbohydrate metabolism of anaerobic fungi. From 10,080 acquired clones, 9,569 clones with average size of 628 bp were selected for analysis. After the assembling process, 1,410 contigs were assembled and 1,369 sequences remained as singletons. 1,192 sequences were matched with proteins in the public data base with known function and 693 of them were matched with proteins isolated from fungi. One hundred and fifty four sequences were classified as genes related with biological process and 328 sequences were classified as genes related with cellular components. Most of the enzymes in the pathway of glucose metabolism were successfully isolated via construction of 10,080 ESTs. Four kinds of hemi-cellulase were isolated such as mannanase, xylose isomerase, xylan esterase, and xylanase. Five $\beta$-glucosidases with at least three different conserved domain structures were isolated. Ten cellulases with at least five different conserved domain structures were isolated. This is the first solid data supporting the expression of a multiple enzyme system in the fungus N. frontalis for polysaccharide hydrolysis.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1457-1463
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• 2005

In this research, the possibilities of using canola meal (CM) in place of soybean meal (SBM), and also the effects of multi-enzyme and phytase supplementation on the performance of quails were investigated. For this purpose, soybean meal (44% CP), canola meal (37% CP), phytase (produced from Peniophora luci) and multi-enzyme ($\beta$-glucanases, pectinases, cellulases and hemicellulases) were used. CM was used supplying 0, 25 and 50% of CP from SBM and each of the phytase and multi-enzyme blends were added to the each level. This study was conducted with 675 day old quails (Coturnix coturnix Japonica) in 9 groups with 3 replicates including 25 birds (mixed sex) per replicate. Nine isocalaric and isonitrogenous diets were prepared. The effects of enzymes and CM levels were studied with a 3${\times}$3${\times}$3 factorial arrangement for three CM levels (0, 25 and 50%), three treatments (without enzyme, phytase enzyme and multi-enzyme) and three replicates. While the 25% CM level did not affect the liveweight gain 50% CM level decreased the liveweight gain (p<0.05). Multi-enzyme addition to the 50% CM group increased the liveweight gain compared to the other groups (p<0.05). CM levels and enzyme supplementation had no effect on feed consumption, feed conversion ratio, dressing percentage, viability, tibia ash content, Ca and P contents of tibia ash, viscera weight, gizzard weight and length of growth period. While heart weight and liver weight were not affected by CM levels, but they were affected by enzyme supplementation. CM levels and enzyme supplementation did not affect final liveweight, feed consumption, feed conversion ratio, egg yield, egg weight, shell weight and shell index during laying period. The increase in the CM level lightened the colour of the yolk (p<0.05).

• Asian-Australasian Journal of Animal Sciences
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• 제31권11호
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• pp.1747-1755
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• 2018

Objective: Common reed (Phragmites communis Trin.) could potentially provide an alternative resource for silage; however, its silage quality is poor. The aim of this study was to investigate the factors in reed that contribute to poor quality and determine how the use of additives at ensiling could improve fermentation quality. Methods: In Experiment 1, we determined the chemical composition and the presence of indigenous lactic acid bacteria (LAB) in reed. We further examined fermentation quality of reed silage under conditions without additives (NA) and treated glucose (G), lactic acid bacteria (L), and their combination (G+L). In Experiment 2, silage of NA, and with an addition of cellulase and lactic acid bacteria (CL) were prepared from harvested reed. The harvested reeds were fertilized at nitrogen concentrations of 0, 4, 8, and $12g\;N/m^2$ and were harvested thrice within one year. Results: The indigenous LAB and fermentable carbohydrates are at extremely low concentrations in reed. Reed silage, to which we added G+L, provided the highest quality silage among treatments in Experiment 1. In Experiment 2, N fertilization had no negative effect on silage quality of reed. The harvest times decreased fermentable carbohydrate content in reed. The CL treatment provided a higher lactic acid content compared to the NA treatment. However, the quality of CL treated silage at the second and third harvests was significantly lower than at the first harvest, due to a reduction in carbohydrates caused by frequent harvesting. Conclusion: The causes of poor quality in reed silage are its lack of indigenous LAB and fermentable carbohydrates and its high moisture content. In addition, reed managed by frequent harvesting reduces carbohydrate content. Although the silage quality could be improved by adding CL, higher-quality silage could be prepared by adding fermentable carbohydrates, such as glucose (rather than adding cellulases).

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.449-452
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• 2002

Trichoderma sp. FJ1의 cellulase 생산을 위한 배지조성의 검토에서, 질소원으로서 0.1% bacto peptone을 사용하였을 때 생산성이 향상되었으며, 순수 상업용기질인 avicel과 섬유소 폐기물인 볏짚의 혼합배양에서 높은 효소생산성이 얻어졌다. 경제적인 효소생산성을 위한 기질로 섬유소 폐기물의 농도와 혼합비를 검토한 결과, 1%농도에서 볏짚과 펄프를 50:50으로 사용했을시 CMCase, xylanase, ${\beta}-glucosidase$, avicelase는 24.3, 38.7, 1.5, 0.6 U/ml가 얻어졌다. 이러한 효소생산성은 타 보고서의 결과보다 동등이상의 우위를 보여주고 있으며 섬유소 폐기물의 생물학적 당화기술에 크게 기여하리라 사료된다.

• 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.65-74
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• 1976

Atternaria sp. was isolated from soil and crude cellulases were prepared from wheat bran culture of the fungus. The activities of the crude enzyme were studied on five different subvstrates and some phsical properties were also examined, crude enzymes were purified by column chromatography on DEAE Sephadex and Sephadex, Isozymes were separated some of which were active specifically on DEAE cellulose and some were primarily active on cellulose and CM-cellulose. The optimal points of pH and temperature for the crude enzyme were varied depending on the substrates ; On cellulose they were at pH 6.0 and 40.deg.C, on CM-cellulose at pH's 4.0 and 6.0 and 60.deg.C, and on DEAW-cellulose at pH 5.0 and 50.deg.C. Two active fractions, F-1 and F-II on Na-CMC was used as substrate the Km values of crude enzyme, F-I and F-II were calculated to be $4{\times}10^{-5}$ , 1.1 * 10$^{-4}$ , and $1.25{\times}10^{-4}mN$ resepctively. The Ki value of $Cu^{++}$ for crude enzyme was$4{\times}^{-4}mN$ , while that of $Nm^{++}$ while in the same concentration of $Mn^{++}$ it reached to 91%. Some 57% activity of F-1 was inhibited in s mN $Cu^{++}$, whereas it was inhibited as much as 81% in the same concentration above the concentration of 0.3 mM with tis activity reaching up to 137% in 2 mM. On the other hand the F-11 was inhibited by the presence of M $n^{++}$ and some 67% activity was inhibited at 2mM.

• 한국균학회지
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• 제14권1호
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• pp.37-41
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• 1986

Pleurotus spp.중 섬유소(纖維素) 분해효소(分解酵素) 생산력(生産力)이 가장 강한 Pleurotus sajor-caju JAFM 1017을 합성배지(合成培地) 상에서 vitamin류(類), 무기염류(無機鹽類)와 배양조건(培養條件)의 영향을 검토(檢討)한 결과(結果)는 다음과 같다. 섬유소(纖維素) 분해효소(分解酵素) 생산(生産)은 folic acid와 thiamine-HCI에 의해 촉진되었고 $KH_2PO_4$와 $MgSO_4$의 최적농도(最適濃度)는 각각 0.2%, 0.04%(w/v)이었으며 그 밖의 무기염류(無機鹽類)는 효과(效果)가 없었다. 효소생산(酵素生産)에 최적(最適)인 배양온도(培養溫度)와 배지(培地) pH는 avicelase가 $25^{\circ}C$, 5.5이었고, CMCase는 $30^{\circ}C$, 5.0이 였으며, ${\beta}-glucosidase$는 $30^{\circ}C$, 6.5이었다.

• Mycobiology
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• 제39권2호
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• pp.103-108
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• 2011

Amylases and cellulases are important enzymes that can be utilized for various biological activities. Ten different wild Nigerian mushrooms (Agaricus blazei, Agaricus sp., Corilopsis occidentalis, Coriolus versicolor, Termitomyces clypeatus, Termitomyces globulus, Pleurotus tuber-regium, Podoscypha bolleana, Pogonomyces hydnoides, and Nothopanus hygrophanus) were assayed for production of these secondary metabolites. The results revealed that most of the tested wild fungi demonstrated very good amylase and cellulase activities. With the incorporation of carboxymethyl-cellulose (a carbon source) into the culture medium, Agaricus blazei had the highest amylolytic activity of 0.60 unit/mL (at $25^{\circ}C$, pH 6.8). This was followed in order by P. tuber-regium and Agaricus sp. with 0.42 and 0.39 unit/mL, respectively ($p {\leq} 0.05$). Maltose and sucrose supplementation into the submerged liquid medium made N. hygrophanus and P. hydnoides to exhibit very low amylase activities of 0.09 and 0.11 unit/mL, respectively. Introducing peptone (an organic nitrogen source) into the basal medium enhanced the ability of C. versicolor to produce a cellulase value of 0.74 unit/mL. Other organic nitrogen sources that supported good cellulase activities were yeast extract and urea. Sodium nitrate (inorganic nitrogen source) generally inhibited cellulase production in all mushrooms. The best carbon source was carboxymethyl-cellulose, which promoted very high cellulase activity of 0.67 unit/mL in C. versicolor, which was followed in order by P. tuber-regium, T. chypeatus, and C. occidentalis ($p {\leq} 0.05$). Sucrose was the poorest carbon compound, supporting the lowest values of 0.01, 0.01, and 0.14 unit/mL in P. hydnoides, A. blazei, and Agaricus sp., respectively.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2014년도 추계학술대회 및 정기총회
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• pp.19-19
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• 2014

Pleurotus ostreatus, P. eryngii, and Flammulina velutipes are major edible mushrooms that account for over 89% of total mushroom production in Korea. Recently, Agrocybe cylindracea, Hypsizygus marmoreus, and Hericium erinaceu are increasingly being cultivated in mushroom farms. In Korea, the production of edible mushrooms was estimated to be 614,224 ton in 2013. Generally, about 5 kg of mushroom substrate is needed to produce 1 kg of mushroom, and consequently about 25 million tons of spent mushroom substrate (SMS) is produced each year in Korea. Because this massive amount of SMC is unsuitable for reuse in mushroom production, it is either used as garden fertilizer or deposited in landfills, which pollutes the environment. It is reasonably assumed that SMS includes different secondary metabolites and extracellular enzymes produced from mycelia on substrate. Three major groups of enzymes such as cellulases, xylanases, and lignin degrading enzymes are involved in breaking down mushroom substrates. Cellulase and xylanase have been used as the industrial enzymes involving the saccharification of biomass to produce biofuel. In addition, lignin degrading enzymes such as laccases have been used to decolorize the industrial synthetic dyes and remove environmental pollutions such as phenolic compounds. Basidiomycetes produce a large number of biologically active compounds that show antibacterial, antifungal, antiviral, cytotoxic or hallucinogenic activities. However, most previous researches have focused on therapeutics and less on the control of plant diseases. SMS can be considered as an easily available source of active compounds to protect plants from fungal and bacterial infections, helping alleviate the waste disposal problem in the mushroom industry and creating an environmentally friendly method to reduce plant pathogens. We describe extraction of lignocellulytic enzymes and antimicrobial substance from SMSs of different edible mushrooms and their potential applications.