• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1110-1117
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• 2001

Responses of the rumen fungus, Neocallimastix frontalis RE1, to long chain fatty acid (LCFA) were evaluated by measuring gas production, filter paper (FP) cellulose digestion and polysaccharidase enzyme activities. LCFA (stearic acid, $C_{18:0}$; oleic acid, $C_{18:1}$; linoleic acid, $C_{18:2}$ and linolenic acid, $C_{18:3}$) were emulsitied by ultrasonication under anaerobic condition, and added to the medium. When N frontalis RE1 was grown in culture with stearic, oleic and linoleic acid, the cumulative gas production, gas pool size, FP cellulose digestion and enzymes activities significantly (p<0.05) increased at some incubation times(especially, exponential phases of fungal growth, 48~120 h of incubation) relative to that for control cultures. However, the addition of linolenic acid strongly inhibited all of the investigated parameters up to 120 h incubation, but not after 168 and 216 h of incubation. These results indicated that stearic, oleic and linoleic acids tended to have great stimulatory effects on fungal cellulolysis, whereas linolenic acid caused a significant (p<0.05) inhibitory effects on the cellulolysis by the rumen fungus. These results are the first report of the effect of LCFAs on the ruminal fungi. Further research is needed to identify the mode of action of LCFAs on fungal strains and to verify whether or not ruminal fungi have ability to hydrate unsaturated LCFAs to saturated FAs. There was high correlation between cumulative in vitro gas production and fungal growth (94.78%), FP cellulose degradation (96.34%), CMCase activity(90.86%) or xylanase activity (87.67%). Thus measuring of cumulative gas production could be a useful tool for evaluating fungal growth and/or enzyme production by ruminal fungi.

• 한국섬유공학회:학술대회논문집
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• 한국섬유공학회 2000년도 봄 한국섬유공학회 학술발표회 논문집(Proceedings of the Korean Textile Conference)
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• pp.125-128
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• 2000

Chemical cellulose from an ascidian tunic is isolated by extraction, digestion and bleaching steps. The content of $\alpha$-cellulose was above 98 wt%, and it's DPw was about 918. A new regenerated cellulose fiber from the chemical cellulose obtained in this study was made using NMMO/water(87/13 wt%) as a solvent by dry jet-wet spinning. The effects of spinning speed and cellulose content of spinning dope on the properties were investigated.

• Asian-Australasian Journal of Animal Sciences
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• 제5권4호
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• pp.635-641
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• 1992

The influence of alkaline hydrogen peroxide (AHP) and peracetic acid treatment on in sacco digestion of aspen was evaluated in three non-lactating ruminally cannulated Holstein cows fed a diet containing 90% forage and 10% concentrate on a DM basis. AHP treatment decreased lignin concentration by 40 to 60% resulting in increased concentrations of neutral detergent fiber (NDF), acid detergent fiber (ADF) and cellulose. Lignin concentrations in peracetic acid treated samples were less than 10% of values for control samples. In sacco disappearance rates of aspen DM, NDF, ADF and cellulose increased (p<.05) with AHP and peracetic acid treatment. Effective degradability of DM, NDF, ADF and cellulose were determined at a ruminal outflow rate of $.05h^{-1}$. Effective degradabilities of AHP treated aspen were approximately three-fold greater and peracetic acid treated samples five-fold greater than untreated control samples. For all parameters measured, peracetic acid treatment resulted in higher (p<.05) digestion coefficients than AHP treated aspen. Results demonstrate that peracetic acid or AHP treatment can enhance the nutritive value of aspen sawdust for ruminants to a level comparable to that reported for many forages.

• Asian-Australasian Journal of Animal Sciences
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• 제22권2호
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• pp.201-205
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• 2009

Effects of Cordyceps militaris mycelia on rumen microbial fermentation were determined by measuring in vitro gas production, cellulose digestion and VFA concentrations. C. militaris mycelia was added to buffered rumen fluid with final concentrations of 0.00, 0.10, 0.15, 0.20, 0.25 and 0.30 g/L and incubation times were for 3, 6, 9, 12, 24, 36, 48 and 72 h. At all incubation times, the gas production showed a quadratic increase with the supplementation of C. militaris mycelia; maximum responses were seen with 0.25 g/L supplementation. However, the gas production was significantly lower for the 0.30 g/L supplementation than for the 0.25 g/L supplementation from 9 h to 72 h incubation. The cellulose filter paper (FP) digestion showed a quadratic increase, as did the gas production except at 3 h incubation. The concentration of total VFA was significantly increased by the supplementation of C. militaris mycelia compared with the control treatment; the highest response was also seen with 0.25 g/L supplementation. This was true for responses in the concentration of acetic and propionic acids. As opposed to other responses, the responses of pH to the supplementation of C. militaris mycelia showed a quadratic decrease from 3 h to 36 h incubation. In conclusion, C. militaris mycelia alter the mixed rumen microbial fermentation with increases in the production of gas and VFA, and cellulose FP digestion.

• Journal of Animal Science and Technology
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• 제47권4호
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• pp.615-624
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• 2005

본 연구는 배양초기 pH 조건이 F. succino- genes의 섬유소 부착과 섬유소 소화에 미치는 영향을 보고자 실시하였다. 선정된 specific primer를 이용하여 F. succinogenes의 genomic DNA로부터 445bp의 16S rDNA 절편을 증폭하여 205bp의 internal control을 제작하였고, cPCR 결과로부터 박테리아 수를 계산할 수 있는 표준곡선의 회귀식($r^2$>0.99)을 얻을 수 있었다. In vitro 배양초기 pH에 따른 F. succ- inogenes의 cellulose 부착을 cPCR로 모니터링한 결과, 발효과정 전 기간동안 초기 pH가 6.8과 6.2일 때 cellulose 건물 g당 부착 균주의 수가 pH 5.6일 때 보다 높았으나, pH 6.8과 6.2 사이에서는 큰 차이는 없었다(p>0.05). Cellulose 분해는 배양시간이 진행됨에 따라 증가 되었으며, 분해 정도는 pH가 증가함에 따라 더 높았다. 배양초기 pH가 6.8, 6.2 그리고 5.8일 때 48시간동안 감소한 pH는 각각 0.24, 0.58 그리고 0.16 이었다. 가스 생산량은 발효 시간이 경과함에 따라 pH가 높을수록 더 많았다. 결론적으로 발효 초기 pH는 F. succinogenes에 의한 cellulose 소화, 가스 생산, pH 변화 및 cellulose 부착에 큰 영향을 주었으며, 특히, 낮은 pH(5.8)에서는 섬유소 소화 및 박테리아 부착을 현저한 감소 시켰다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1164-1173
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• 2006

Cellulase from Aspergillus niger, (${\alpha}$-amylase from Bacillus sp. and protease from Bacillus globigii were used as enzyme sources in this study to examine how their respective substrates protect them in two kinds of simulated gastrointestinal tract digesting processes. Avian total digest tract simulation test showed that filter paper, Avicel and cellulose resulted in 7.7, 6.4 and 7.4 times more activity than of unprotected cellulose, respectively. Protease with addition of casein, gelatin or soybean protein showed no significant protection response. Starch protected amylase to be 2.5 times activity of the unprotected one. Monogastric animal total tract digestion simulation test showed that filter paper, Avicel and cellulose resulted in 5.9, 9.0 and 8.8 times activity of unprotected cellulase, respectively. Casein, gelatin and soybean protein resulted in 1.2, 1.3 and 2.0 times activity of unprotected protease, respectively. Starch did not protect amylase activity in monogastric animal total tract simulation. Protection of mixed enzymes by substrates in two animal total tract simulation tests showed that filter paper in combination with soybean protein resulted in 1.5 times activity of unprotected cellulose, but all substrates tested showed no significant protection effect to protease. Soybean protein and starch added at the same time protected the amylase activity to be two times of the unprotected one. Test of non-purified substrate protection in two animal total digest tract simulation showed that cellulase activity increased as BSA (bovine serum albumin) concentration increased, with the highest activity to be 1.3 times of unprotected enzyme. However, BSA showed no significant protection effect to protease. Amylase activity increased to 1.5 times as BSA added more than 1.5% (w/v). Cellulase activity increased to 1.5 times as soybean hull was added higher than 1.5%. Amylase had a significant protection response only when soybean hull added up to 2%. Protease activity was not protected by soybean hull to any significant extent.

• Asian-Australasian Journal of Animal Sciences
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• 제22권1호
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• pp.131-138
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• 2009

Among rumen microbes, bacteria play important roles in the biological degradation of plant fiber due to their large biomass and high activity. To maximize the utilization of fiber components such as cellulose and hemicellulose by ruminant animals, the ecology and functions of rumen bacteria should be understood in detail. Recent genome sequencing analyses of representative fibrolytic bacterial species revealed that the number and variety of enzymes for plant fiber digestion clearly differ between Fibrobacter succinogenes and Ruminococcus flavefaciens. Therefore, the mechanism of plant fiber digestion is also thought to differ between these two species. Ecology of individual fibrolytic bacterial species has been investigated using pure cultures and electron microscopy. Recent advances in molecular biology techniques complement the disadvantages of conventional techniques and allow accurate evaluation of the ecology of specific bacteria in mixed culture, even in situ and in vivo. Molecular monitoring of fibrolytic bacterial species in the rumen indicated the predominance of F. succinogenes. Nutritive interactions between fibrolytic and non-fibrolytic bacteria are important in maintaining and promoting fibrolytic activity, mainly in terms of crossfeeding of metabolites. Recent 16S rDNA-based analyses suggest that presently recognized fibrolytic species such as F. succinogenes and two Ruminococcus species with fibrolytic activity may represent only a small proportion of the total fibrolytic population and that uncultured bacteria may be responsible for fiber digestion in the rumen. Therefore, characterization of these unidentified bacteria is important to fully understand the physiology and ecology of fiber digestion. To achieve this, a combination of conventional and modern techniques could be useful.

• 한국폐기물자원순환학회지
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• 제34권5호
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• pp.474-481
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• 2017

This study was conducted to investigate the effects of inoculum and carbon sources on anaerobic digestion characteristics. The treatments were combinations of inoculum (digestate of cattle manure and rumen fluid) with carbon sources (starch, cellulose, and xylan). Anaerobic digestion was performed in triplicate at $37^{\circ}C$ for 18 days at 100 rpm. Sampling was performed at 0, 1, 2, 3, 4, 5, 7, 9, 12, 15, and 18 days to measure pH, ammonia-N, volatile solids reduction, the cumulative methane content, and the cumulative methane production. There was a significant difference in methane content depending on the carbon source and there was a significant difference in pH, ammonia-N, methane production, and methane content depending on the inoculum (P < 0.05). The results of methane production were higher in the digestate of cattle manure treatment than in the rumen fluid treatment (P < 0.05). In this study, different digestive patterns depending on the type of carbon source could be used as basic research data to set the hydraulic residence time of anaerobic digestion facilities. In addition, the use of ruminal fluid as an inoculum may help accelerate the hydrolysis and acid production steps.

• 미생물학회지
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• 제25권4호
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• pp.297-297
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• 1987

A $C_{1}$ enriched cellulase fraction was separated from culture filtrate of anaerobic Clostridium thermocellum by hydroxyapatite column chromatography. The separated fraction showed strong synergistic action with $C_{x}$ component (endo-$\beta$-1, 4-glucanase) in digestion of crystalline cellulose, similar to the other aerobic cellulolytic microorganisms. Unlike the $C_{x}$ component the $C_{1}$ enriched fraction was rapidly inactivated by oxidation at the atmospheric condition. The enzyme activity was significantly enhanced by the addition of reducing agents, especially $\beta$-mercaptoethanol, which indicates that a $C_{1}$ component has a lot of sulfhydryl groups essential for the enzyme activity. The effect of metal ions on $C_{1}$ activity was also investigated. The $C_{1}$ fraction was found to be thermally stable compare to endo-$\beta$-1,4-glucanase. Optimal temperature and pH were found to be 60.deg.C and 6.0, respectively.

• Applied Biological Chemistry
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• 제21권2호
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• pp.112-118
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• 1978

소의 갑상선에서 추출한 Xanthine oxidase를 disc gel eleectrophoresis로서 정제도(Purity)를 측정하여 Xanthine oxidase 이외의 다른 불순 단백질이 나타나지 않을 때까지 정제하였다. 그 정제 과정은 Pancreatin digestion, butanol 추출, ammonium sulfate 단백질 침전, calcium phosphate gel-cellulose column chromatography, gel filtration, preparative Sephadex G-25 column electrophoresis와 Preparative polyacrylamide gel electrophoresis 등을 포함하고 있다. 이러한 과정을 통하여 갑상선 Xanthine oxidase는 1,000배 정도 정제되었다. 그러나 효소의 비활성도(Specific activity)는 우유에서 추출한 이 효소에 상응하는 정도로 정제된 효소의 비활성도와 비교 되었을 때 지극히 낮았다. 갑상선 Xanthine oxidase도 효소 반응에 필요한 기질과 electron acceptor의 특수성(Specificity)이 어느 특수한 한 기질에 한정되지 않았음을 보였고 Kinetic 성질도 우유에서 추출된 Xanthine oxidase와 비교하였을 때 가장 일반적인 Xanthine oxidase 기질에 대한 Michaelis 상수(Km)가 약간의 예외도 있었으나 상당히 비슷하였다.