• Journal of Microbiology
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• 제42권3호
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• pp.205-210
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• 2004

The sequence coding for carboxymethylcellulase (CMCase, CelC) was isolated from the DNA of Salmonella typhimurium URl. Comparison between the deduced amino acid sequence of CelC (368 amino acid residues, Molecular mass 41 kDa) and that of the previously published CMCase revealed that this enzyme belongs to the cellulase family 8 and D. The protein was overproduced in Escherichia coli using T7 expression system, and its activity was confirmed by CMC-SDS-PAGE. When the overexpressed CelC protein was tested on cellulose-type substrates, the recombinant protein is able to degrade cellulose-type substrates, such as CM-cellulose, xylan, avicel, lichenan, and laminarin. Optimal temperature and pH for enzyme activity were found to be 50$^{\circ}C$ and pH 6.5, respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제17권2호
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• pp.199-202
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• 2004

Cellulolytic bacterial strains have been isolated from the faeces of wild (blackbuck, Antilope cervicapra; nilgai, Baselophus tragocamelus chinkara, Gazella gazella spotted deer, Axis axis and hog deer, Cervus porcinus) and rumen liquor of domestic (sheep, Ovis aries) ruminants. Five best cellulose degrading bacterial isolates (Ruminococcus sp.) were used as microbial feed additive along with buffalo rumen liquor as inoculum to study their effect on digestibility of feed and enzyme production in in vitro conditions. The bacterial isolate from chinkara (CHI-2) showed the highest per cent apparent dry matter (DM) digestibility ($35.40{\pm}0.60$), true dry matter digestibility ($40.80{\pm}0.69$) and NDF ($26.38{\pm}0.83$) digestibility (p<0.05) compared to control ($32.73{\pm}0.56$, $36.64{\pm}0.71$ and $21.16{\pm}0.89$, respectively) and other isolates at 24 h of incubation with lignocellulosic feeds (wheat straw and wheat bran, 80:20). The same isolate also exhibited the highest activities of fibre degrading enzymes like carboxymethylcellulase, xylanase, ${\beta}$-glucosidase and acetyl esterase. The bacterial isolate from chinkara (Gazella gazella) appears to have a potential to be used as feed additive in the diet of ruminants for improving utilization of nutrients from lignocellulosic feeds.

• 한국초지조사료학회지
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• 제11권1호
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• pp.43-52
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• 1991

The overall objective of this research was to evaluate the effects of spraying sulfuric acid solution, and adding corn meal or molasses to reed canarygrass silage on digestibility of dry matter (DM) and fibrous components in ruminant. DM content of molasses or corn meal treated silages were higher than control. Whereas, sulfuric acid treated silage showed lower DM content compared with untreated control. Acid detergent fiber content was similar among treatments, but neutral detergent fiber (NDF) and hemicellulose concentration were higher in treated silages than those of control. Organic acids content were also increased in treated silages. In contrast, silage pH were lower in treated silages. Treated silages increased both rate and extent of DM and NDF in situ digestibility compared to control at all incubation times. Especially, 7 % molasses and 0.4 % sulfuric acid treated silages increased DM and NDF digestibility significantly after 12 hours of incubation compared with control. In nitro DM and NDF digestibility showed similar trend to in situ incubation. Also treated silages(mo1asses or sulfuric acid treated) incubated in situ had higher particle-associated carboxymethylcellulase activity throughout the incubation except 72 hours.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.518-521
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• 1999

Bacillus sp. 79-23 spores were irradiated with $^{60}Co$ gamma-rays at doses ranging from 0.5 to 5 kGy. Following gamma-irradiation, seven mutant strains were isolated by scoring the halo sizes formed around the colonies grown on LB agar plates containing 4% carboxymethylcellulose (CMC) and trypan blue. The mutant strains showed a 1.5 to 2-fold increase in carboxymethylcellulase (CMCase) activity over the parent strain. Wheat bran acted as an effective inducer for CMCase production in the parent and mutant strains. Mutant strains 68 and 70 were identified as exhibiting higher CMCase activities than those of other mutants in LB media both with and without 3% wheat bran. In addition, these strains seem to produce substantially lower amounts of capsular materials, whereas the parent strain produced large amounts of them in both liquid and solid LB media. In flask cultures, the CMCase production by mutants 68 and 70 reached maximum levels of 17.5 unit/ml and 15.7 unit/ml, respectively, in an LB medium containing 3% wheat bran.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권1호
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• pp.44-47
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• 1998

Maximum cellulase production was sought by comparing the activities of the cellulases produced by different Trichoderma reesei strains and Aspergillus niger. Trichoderma reesei Rut-C30 showed higher cellulase activity than other Trichoderma reesei stains and Aspergillus niger that was isolated from soil. By optimizing the cultivation conditions during shake flask culture, higher cellulase production could be achieved. The FP(filter paper) activity of 3.7U/ml and CMCase (Carboxymethylcellulase) activity of 60U/ml were obtained from shake flask culture. When it was grown in 2.5L fermentor, where pH and DO levels are controlled, the enzyme activities were 133.35U/ml (CMCase) and 11.67U/ml(FP), respectively. Ammonium sulfate precipitation method was used to recover enzymes from fermentation broth. The dried cellulase powder showed 3074.9U/g of CMCase activity and 166.7U/g of FP activity with 83.5% CMCase recovery.

• 한국균학회지
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• 제43권3호
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• pp.165-173
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• 2015

감자덩이버섯(Mattirolomyces terfezioides)은 단맛을 가진 트러플의 한 종류로, 주로 아까시나무(Robinia pseudoacacia)와 외생균근을 형성한다. 상업적으로 인공재배할 만한 가치가 있는 버섯으로, 본 연구에서는 우리나라 아까시나무 숲에서 수집한 감자덩이버섯균의 균사 생장특성과 효소활성에 대해 조사한 바를 처음으로 보고하고자 한다. 이 균은 PDA 배지, $30^{\circ}C$에서 가장 잘 생장하였고, 액체배양 시에는 pH 8.0의 MMN 배지에서 균 생장력이 가장 우수하였다. 또한 암모늄염보다는 질산염 형태의 무기질소원이 함유된 액체배지에서 균체량이 1.8배 더 높았으며, 효소 검색용 고체배지 상에서 carboxymethylcellase와 laccase 활성을 나타냈다.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1280-1290
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• 2014

In order to obtain the cellulolytic bacterial consortia, sediments from Great Basin hot springs (Nevada, USA) were sampled and enriched with cellulosic biomass as the sole carbon source. The bacterial composition of the resulting anaerobic ethanol-producing celluloytic bacterial consortium, named SV79, was analyzed. With methods of the full-length 16S rRNA library-based analysis and denaturing gradient gel electrophoresis, 21 bacteria belonging to eight genera were detected from this consortium. Clones with closest relation to the genera Acetivibrio, Clostridium, Cellulosilyticum, Ruminococcus, and Sporomusa were predominant. The cellulase activities and ethanol productions of consortium SV79 using different agricultural residues (sugarcane bagasse and spent mushroom substrate) and energy crops (Spartina anglica, Miscanthus floridulus, and Pennisetum sinese Roxb) were studied. During cultivation, consortium SV79 produced the maximum filter paper activity (FPase, 9.41 U/ml), carboxymethylcellulase activity (CMCase, 6.35 U/ml), and xylanase activity (4.28 U/ml) with sugarcane bagasse, spent mushroom substrate, and S. anglica, respectively. The ethanol production using M. floridulus as substrate was up to 2.63 mM ethanol/g using gas chromatography analysis. It has high potential to be a new candidate for producing ethanol with cellulosic biomass under anoxic conditions in natural environments.

• Asian-Australasian Journal of Animal Sciences
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• 제16권3호
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• pp.374-379
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• 2003

Four rumen fistulated Murrah buffaloes were used to study the effect of four diets differing in roughage to concentrate ratio on rumen biochemical changes, microbial enzyme profile and in sacco degradability of feed in a $4{\times}4$ Latin Square design. The animals were fed four diets consisting of 80:20, 70:30, 60:40 and 50:50 ratios of wheat straw and concentrate mixtures, respectively. Wheat straw and concentrate mixture were mixed with water (0.6 l/kg feed) and complete feed mixture was offered to the animals at 8:00 h and 16:00 h in two equal parts. The variation in pH of rumen liquor (difference of maximum and minimum during 0-8 h post feeding) increased with increasing level of concentrate mixture in the diet. There was no effect of diet composition on volatile fatty acids, total nitrogen and trichloro-acetic acid precipitable nitrogen in the rumen liquor, but ammonia nitrogen increased with increasing level of concentrate mixture in the ration. Major portions of all fibre degrading enzymes were present in the particulate material (PM) of the rumen contents, but protease was absent in PM fraction. The activities of micro-crystalline cellulase, acetyl esterase and protease increased with increase in the level of concentrate mixture, but the activities of other enzymes (carboxymethylcellulase, filter paper degrading activity, xylanase, $\beta$-glucosidase and $\beta$-xylosidase) were not affected. The in sacco degradability and effective degradability of feeds increased with increasing level of concentrate mixture in the ration.

• Asian-Australasian Journal of Animal Sciences
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• 제18권8호
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• pp.1110-1115
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• 2005

To study the effect of feeding lactic acid producing bacteria on the performance of cattle calves, twenty four, day old male crossbred cattle calves (Bos indicus${\times}$Bos taurus), were distributed into two groups of 12 animals each. The animals were fed on calf starter containing wheat bran and green berseem ad libitum and milk as per requirement upto 8 weeks of age. The diet of calves of Group 2 was supplemented with 500 ml culture of Lactobacillus acidophilus-15. Total duration of the experiment was 31 weeks. There was no significant difference in intake and digestibility of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF) and crude protein (CP) between the groups. The rumen pH, protozoa numbers, concentration of volatile fatty acids (VFA), ammonia nitrogen ($NH_3-N$), trichloroacetic acid precipitable nitrogen (TCA-ppt N) and activity of microbial enzymes (carboxymethylcellulase, xylanase, amylase and protease) were not affected due to probiotic supplementation. Average live weight gain of the calves was improved (about 10%) and feed:gain ratio was reduced (about 5%) in the animals given Lactobacillus culture. The data indicated that crossbred calves could be reared on a diet devoid of cereal grain and addition of Lactobacillus culture in the diet resulted in an added advantage in growth performance of the animals.

• 한국유기농업학회지
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• 제20권3호
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.